Lipidic furan, pyrrole, and thiophene compounds for treatment of cancer, neurological disorders, and fibrotic disorders

ABSTRACT

Compounds, methods, and compositions are provided for the treatment of cancer, neurological disorders, and fibrotic disorders. Specifically, the invention includes administering an effective amount of a compound of Formula I, II, or III, or a pharmaceutically acceptable composition, salt, isotopic analog, prodrug, or combination thereof, to a subject suffering from a cancer, neurological disorder, or fibrotic disorder.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/721,638, filed Dec. 19, 2019, which is a continuation of U.S. patentapplication No. Ser. No. 16/110,778, filed Aug. 23, 2018, which is acontinuation of U.S. patent application Ser. No. 15/782,535, filed Oct.12, 2017, which is a continuation of U.S. patent application Ser. No.15/162,074, filed May 23, 2016, which is a continuation of U.S. patentapplication Ser. No. 14/229,130, filed Mar. 28, 2014, which claims thebenefit of U.S. Provisional Application No. 61/853,163, filed Mar. 29,2013, the contents of which are incorporated by reference in theirentireties.

FIELD OF THE INVENTION

The present invention is directed to compounds, methods, andcompositions for the treatment of cancer, neurological disorders, andfibrotic disorders.

BACKGROUND

Every year, cancer claims the lives of more than half a millionAmericans. Cancer is the second leading cause of death in the UnitedStates, exceeded only by heart disease. One of every four deaths in theUnited States is cancer-related.

Cancers arise from cells that have undergone genetic alterations,leading to abnormal proliferation on a clonal basis. These geneticalterations can include activation of oncogenes or inactivation of tumorsuppressors. Different types of cancers have been found to have a widerange of underlying genetic alterations and vary in their pathologicalprogression to the cancerous state, including in their ability to invadesurrounding normal tissues and metastasize.

Conventional approaches to cancer treatment include surgery, radiation,and chemotherapy, or a combination thereof. However, for more aggressiveand invasive cancers, these treatments have less of an effect thancancers caught at earlier stages of progression. Firstly, invasivecancers appear to be inherently more resistant to a wider variety ofchemotherapeutic agents. Secondly, the invasive progression of andmetastatic potential of cancer is complex and still poorly understood.

Likewise, neurological disorders such as Parkinson's disease,Alzheimer's disease, or other neurodegenerative disorders that generallystart late in life affect nearly one in three seniors. Currently, thereare no cures for these types of disorders, and researchers are lookingfor new treatments to alter the course of the disease and improve thequality of life for those with dementia-like disorders.

Fibrotic diseases, which include pulmonary fibrosis, systemic sclerosis,liver cirrhosis and cardiovascular disease, account for over 45% ofdeaths in the developed world. In the United States, for example, whilesome prescribed medications may stabilize subjects who have pulmonaryfibrosis, there are currently no FDA-approved therapies, and lungtransplantation remains the most viable course of treatment to extendthe lives of those with pulmonary fibrosis.

Accordingly, there is a continuing need to identify new treatments totarget these devastating disorders.

SUMMARY

Compounds, methods, and compositions are provided for the treatment ofcancer, neurological disorders, and fibrotic disorders. Specifically,the invention includes administering an effective amount of a compoundof Formula I, II, or III, or a pharmaceutically acceptable composition,salt, isotopic analog, prodrug, or combination thereof, to a subjectsuffering from a cancer, neurological disorder, or fibrotic disorder,wherein Formula I, II or III are:

wherein;each Z is independently O, S or NR₅;wherein R₁, R₂, R₃ and R₄, are the same or different, separatelyrepresent a hydrogen atom, a C₁-C₃₅ alkyl radical, advantageouslyC₁₀-C₂₂, more advantageously C₁₂-C₂₀, further advantageously C₁₃-C₁₇; aC₁-C₃₅ alkenyl radical, advantageously C₁₀-C₂₂, more advantageouslyC₁₂-C₂₀, further advantageously C₁₃-C₁₇; or a C₁-C₃₅ alkynyl radical,advantageously C₁₀-C₂₂, more advantageously C₁₂-C₂₀, furtheradvantageously C₁₃-C₁₇, and wherein the alkyl, alkenyl or alkynyl moietyis optionally substituted with one or more halogens (F, Cl, Br, or I,and more typically F) and/or by one or more moieties selected from thegroup consisting of epoxide (e.g, an oxygen divalently linked to thecarbon chain), hydroxyl or protected hydroxyl (OR₅), thiol or protectedthiol (SR₅), amine (NR₅R₆), aldehyde (—CHO), ketone (—COR₅), acetyl(—O—CO—R₅), or ester (—C(O)OR₅) function and wherein R₅ and R₆separately represent a hydrogen atom, a C₁-C₃₅, more typically a C₁ toC₂₀ alkyl radical, advantageously C₁₀-C₂₂, more advantageously C₁₂-C₂₀,further advantageously C₁₃-C₁₇, or a C₁-C₃₅ alkenyl radical,advantageously C₁₀-C₂₂, more advantageously C₁₂-C₂₀, or furtheradvantageously C₁₃-C₁₇.

In one embodiment, R₁ is a C₁₂, C₁₃, C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, C₁₉, C₂₀,C₂₁, C₂₂, C₂₃, C₂₄, or C₂₅ saturated carbon chain. In anotherembodiment, R₁ is a C₁₂, C₁₃, C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, C₁₉, C₂₀, C₂₁,C₂₂, C₂₃, C₂₄, or C₂₅ carbon chain optionally with one, two or threedouble or triple bonds or a combination thereof. In one embodiment theR₁ has one, two or three double bonds, and wherein the two double bondscan be conjugated or non-conjugated and wherein the three double bondscan be fully, partially or non-conjugated. In one embodiment, a doublebond can be in the terminal position. In another embodiment, R₁ is aC₁₂, C₁₃, C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, C₁₉, C₂₀, C₂₁, C₂₂, C₂₃, C₂₄, or C₂₅carbon chain with at least one double and at least one triple bond. Inyet another embodiment, R₁ is a C₁₂, C₁₃, C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, C₁₉,C₂₀, C₂₁, C₂₂, C₂₃, C₂₄, or C₂₅ carbon chain with one triple bond. Inone embodiment, the triple bond can be in the terminal position.

In one embodiment, R₁ is a C₁-25 saturated carbon chain. In oneembodiment, R₁ is a C₁₃-C₁₇ saturated carbon chain. In one embodiment,R₁ is a C₁₁, C₁₃, C₁₅, C₁₇, C₁₉, C₂₁, C₂₃, or C₂₅ saturated carbonchain. In another embodiment, R₁ is a C₁₁, C₁₃, C₁₅, C₁₇, C₁₉, C₂₁, C₂₃,or C₂₅ carbon chain optionally with one, two or three double or triplebonds or a combination thereof. In one embodiment the R₁ has one, two orthree double bonds, and wherein the two double bonds can be conjugatedor non-conjugated and wherein the three double bonds can be fully,partially or non-conjugated. In one embodiment, a double bond can be inthe terminal position. In another embodiment, R₁ is a C₁₁, C₁₃, C₁₅,C₁₇, C₁₉, C₂₁, C₂₃, or C₂₅ carbon chain with at least one double and atleast one triple bond. In yet another embodiment, R₁ is a C₁₁, C₁₃, C₁₅,C₁₇, C₁₉, C₂₁, C₂₃, or C₂₅ carbon chain with one triple bond. In oneembodiment, the triple bond can be in the terminal position.

The double bond can be in the Z or E configuration. In one embodiment,R₁ has one or two double bonds in the Z configuration. In an alternativeembodiment, one or two double bonds are in the E configuration. Forexample, when conjugated, the double bonds can be: (2Z, 5Z); (3Z, 6Z),(7Z, 10Z), (8Z, 11Z), (9Z, 12Z), (10Z, 13Z), (11Z, 14Z), or (12Z, 15Z).In an alternative embodiment, R₁ has a single double bond in the Zconfiguration. In one embodiment, the double bond can be (2Z), (3Z),(4Z), (5Z), (6Z), (7Z), (8Z), (9Z), (10Z), (11Z), (12Z), (13Z), (14Z),(15Z), (16Z), (17Z), (18Z), or (19Z). In one embodiment, R₁ has a singledouble bond in the E configuration. In one embodiment, the double bondcan be (2E), (3E), (4E), (5E), (6E), (7E), (8E), (9E), (10E), (11E),(12E), (13E), (14E), (15E), (16E), (17E), (18E), or (19E). In oneembodiment, the double bonds can be: (2Z, 5E); (3Z, 6E), (7Z, 10E), (8Z,11E), (9Z, 12E), (10Z, 13E), (11Z, 14E), or (12Z, 15E). In oneembodiment, the double bonds can be: (2E, 5Z); (3E, 6Z), (7E, 10Z), (8E,11Z), (9E, 12Z), (10E, 13Z), (11E, 14Z), or (12E, 15Z).

In one embodiment, R₁ is substituted with at least one OR₅ group. In onesub-embodiment, R₁ is substituted with at least one OR₅ group whereinR₅=H.

In one embodiment, R₁ is substituted with at least two OR₅ groups. Inone sub-embodiment, R₁ is substituted with at least two OR₅ groupwherein R₅=H.

In one embodiment, R₁ is substituted with at least three OR₅ groups. Inone sub-embodiment, R₁ is substituted with at least three OR₅ groupwherein R₅=H.

In one embodiment, R₁ is substituted with at least one OR₅ group. In onesub-embodiment, R₁ is substituted with at least one OR₅ group whereinR₅=C(O)CH₃.

In one embodiment, R₁ is substituted with at least two OR₅ groups. Inone sub-embodiment, R₁ is substituted with at least two OR₅ groupwherein R₅=C(O)CH₃.

In one embodiment, R₁ is substituted with at least three OR₅ groups. Inone sub-embodiment, R₁ is substituted with at least three OR₅ groupwherein R₅=C(O)CH₃.

In one embodiment, R₁ is substituted with at least one NR₅R₆ group. Inone sub-embodiment, R₁ is substituted with at least one NR₅R₆ groupwherein R₅=R₆=H.

In one embodiment, R₁ is substituted with at least two NR₅R₆ groups. Inone sub-embodiment, R₁ is substituted with at least two NR₅R₆ groupwherein R₅=R₆=H.

In one embodiment, R₁ is substituted with at least three NR₅R₆ groups.In one sub-embodiment, R₁ is substituted with at least three NR₅R₆ groupwherein R₅=R₆=H.

In one embodiment, R₁ is CH₃—(CH₂)_(m)—(CH═CH)_(x)—(CH₂)_(m) wherein n,m and x do not equal 0 and m+2x÷=1 to 35.

In some embodiments, the compound has Formula I:

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a C₉-C₂₀ alkyl chain comprising two or more double bondsoptionally substituted as defined above.

In some embodiments, the compound has Formula II:

wherein;R₁ is a C₉-C₂₀ alkyl chain comprising two or more double bondsoptionally substituted as defined above.

In some embodiments, the compound has Formula III:

wherein;R₁ is a C₉-C₂₀ alkyl chain comprising two or more double bondsoptionally substituted as defined above.

In an alternative embodiment, the furan, thiophene, or pyrrole can befused to another heterocyclic or heteroaromatic moiety to produce amulti-ring core, for example, benzofuran, benzothiophene, or indole,which may optionally be substituted with one or more functional groups,preferably with one or more alkyl, alkoxy, halo, or hydroxysubstituents.

In one embodiment, a compound of Formula I, Formula II or Formula IIIhas a purity of greater than 95%, greater than 96%, greater than 97%,greater than 98%, or greater than 99%.

In one embodiment provided herein is a method of reducing growth of asolid tumor in a subject, comprising administering an effective amountof a compound of Formula I, II, or III, or a pharmaceutically acceptablecomposition, salt, isotopic analog, prodrug, or combination thereof. Thetumor can be a primary tumor or a metastatic tumor. In one embodiment, atumor is, for example, a colon tumor, an ovarian tumor, a lung tumor, anesophageal tumor, a breast tumor, a prostate tumor, a carcinoma, or acancer of the skin. In one embodiment, the compound administered isFormula I(n).

In one embodiment, provided herein is a method for inhibiting thecellular migration of cancer cells in a subject comprising administeringan effective amount of a compound of Formula I, II, or III, or apharmaceutically acceptable composition, salt, isotopic analog, prodrug,or combination thereof. In one embodiment, the compound administered isFormula I(n).

In one embodiment provided herein is a method of treating a skin canceror pre-cancerous lesion of the skin, comprising administering aneffective amount of a compound of Formula I, II, or III, or apharmaceutically acceptable composition, salt, isotopic analog, prodrug,or combination thereof. In one embodiment, the skin cancer orpre-cancerous lesion of the skin is selected from the group consistingof all pre-cancerous lesions, such as actinic keratosis and leukoplakia,and of all non-melanoma skin cancers such as squamous cell carcinoma andbasal cell carcinoma. In one embodiment, the compound administered isFormula I(n).

The compounds described herein can be administered to the subject incombination with other chemotherapeutic agents used for the treatment ofcancer or proliferative disorders. If convenient, the compoundsdescribed herein can be administered at the same time as anotherchemotherapeutic agent, in order to simplify the treatment regimen. Insome embodiments, the compound and the other chemotherapeutic can beprovided in a single formulation. In one embodiment, the use of thecompounds described herein is combined in a therapeutic regime withother agents.

Also provided herein is a method of treating a neurological disorder ina subject comprising administering an effective amount of a compound ofFormula I, II, or III, or a pharmaceutically acceptable composition,salt, isotopic analog, prodrug, or combination thereof. In oneembodiment, the neurological disorder is Alzheimer's disease. In oneembodiment, the neurological disorder is Parkinson's disease. In oneembodiment, the neurological disease is dementia. In one embodiment, thecompound administered is Formula I(n).

In one embodiment provided herein is a method of treating a fibroticdisorder, comprising administering an effective amount of a compound ofFormula I, II, or III, or a pharmaceutically acceptable composition,salt, isotopic analog, prodrug, or combination thereof. In oneembodiment, the fibrotic disorder is selected from rheumatoid arthritis,surgical adhesions, osteoarthritis, visible skin scars, or acardiovascular, liver, kidney, lung or periodontal fibrotic disorder,diseases entailing excess collagen and elastin deposition, cutaneouskeloid formation, progressive systemic sclerosis, liver cirrhosis,idiopathic and pharmacologically induced pulmonary fibrosis, chronicgraft-versus-host disease, scleroderma (local and systemic), Peyronie'sdisease, pharmacologically induced fibrosis of the penis,post-cystoscopic urethral stenosis, post-surgical internal adhesions,myelofibrosis, and idiopathic and pharmacologically inducedretroperitoneal fibrosis. In one embodiment, the fibrotic disorder isrheumatoid arthritis.

In some embodiments, the subject or host is a mammal, including a human.The compound can be administered to the subject by any desired route,including intravenous, sublingual, buccal, oral, intraaortal, topical,intranasal, parenteral, transdermal, systemic, intramuscular, or viainhalation.

In summary, the present invention includes the following features:

A) Compounds of Formula I, II, or III as described herein, andpharmaceutically acceptable compositions, salts, isotopic analogs, orprodrugs thereof, for use in the inhibition of the proliferation ofcancer cells in a subject. In one embodiment, the compound is FormulaI(n);B) Compounds of Formula I, II, or III as described herein, andpharmaceutically acceptable compositions, salts, isotopic analogs, orprodrugs thereof, for use in the inhibition of the migration of cancercells in a subject. In one embodiment, the compound is Formula I(n);C) A compound as described herein, or a pharmaceutically acceptablecomposition, salt, isotopic analog, or prodrug thereof, for use as achemotherapeutic in the treatment of cancer. In one embodiment, thecompound is Formula I(n);D) A compound as described herein, or a pharmaceutically acceptablecomposition, salt, isotopic analog, or prodrug thereof, for use incombination with a second chemotherapeutic agent in a subject undergoinga therapeutic regime to treat a cancer. In one embodiment, the compoundis Formula I(n);E) Use of a compound described herein, or a pharmaceutically acceptablecomposition, salt, isotopic analog, or prodrug thereof, in themanufacture of a medicament for use as a chemotherapeutic to treat asubject with a cancer;F) Processes for the preparation of therapeutic products that contain aneffective amount of a compound described herein, for use in thetreatment of a subject having a cancer;G) A method for manufacturing a medicament selected from the compoundsdescribed herein intended for therapeutic use as a chemotherapeutic forthe treatment of a cancer.H) Compounds of Formula I, II, or III as described herein, andpharmaceutically acceptable compositions, salts, isotopic analogs, orprodrugs thereof, for use in the treatment of a subject suffering from aneurological disorder. In one embodiment, the compound is Formula I(n);I) Use of a compound described herein, or a pharmaceutically acceptablecomposition, salt, isotopic analog, or prodrug thereof, in themanufacture of a medicament for use as a treatment in a subject with aneurological disorder;J) Processes for the preparation of therapeutic products that contain aneffective amount of a compound described herein, for use in thetreatment of a subject having a neurological disorder;K) A method for manufacturing a medicament selected from the compoundsdescribed herein intended for therapeutic use as a in the treatment of aneurological disorder;L) Compounds of Formula I, II, or III as described herein, andpharmaceutically acceptable compositions, salts, isotopic analogs, orprodrugs thereof, for use in the treatment of a subject suffering from afibrotic disorder. In one embodiment, the compound is Formula I(n);M) Use of a compound described herein, or a pharmaceutically acceptablecomposition, salt, isotopic analog, or prodrug thereof, in themanufacture of a medicament for use as a treatment in a subject with afibrotic disorder;N) Processes for the preparation of therapeutic products that contain aneffective amount of a compound described herein, for use in thetreatment of a subject having a fibrotic disorder;O) A method for manufacturing a medicament selected from the compoundsdescribed herein intended for therapeutic use as a in the treatment of afibrotic disorder.P) Compounds as described herein for use to treat a solid tumor,neurological disorder, or fibrotic disorder, wherein the activeingredient is not provided as a botanical extract mixture orcombination, but instead the active compound is delivered in a highlypure form. In one embodiment, the invention is a dosage form for thetreatment of a solid tumor, neurological disorder, or fibrotic disorder,wherein the active compound has a purity of at least 96%, 97%, 98%, or99%, without respect to fillers, stabilizers, or other inert or inactiveingredients. In an alternative embodiment, the dosage form has two ormore active ingredients, wherein only one of the active ingredients isselected from compounds of Formula I, II or III as described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is graph of cell migration using the breast cancer cell lineMDA-MB231 for cells treated with 0 μl/10 ml, 5 μl/10 ml and 10 μl/10 mlFormula I(n). The effect of Formula I(n) on the migration of MDA-MB231invasive breast tumor cells was tested using the Membrane InvasionCulture System (MICS) chamber. The Membrane Invasion Culture System(MICS) chamber was assembled with Crosstex 10 μm polycarbonate membranepre-soaked in gelatin for 12 hours. The lower wells were fully loadedwith DMEM supplemented with 10% FBS and 0, 5, 10 μl/10 ml Formula I(n)diluted in 0-20 μl/10 ml methanol. The upper wells were loaded with 1 mlserum-free phenol-red-free media and corresponding Formula I(n)/methanolsolution. 100,000 cells/well were loaded and the chamber was incubatedat 37° C. for 24 hours. After the incubation the media from the upperpart of the chamber was removed and non-migratory cells from the uppersurface of the membrane were wiped away with Kimwipe. The cells werefixed onto the membrane by immersing it in 100% MeOH. Cells were stainedin Eosin solution for 25 seconds and in crystal violet solution for 35sec. The membrane was rinsed in water, placed onto a microscope slidepre-treated with immersion oil, covered with a coverslip and cells werecounted under a light microscope. Each sample was measured intriplicate. As described in Example 6, Formula I(n) significantlyinhibited MDA-MB231 tumor cell migration. FIG. 1B is graph of cellmigration using the breast cancer cell line Hs578T for cells treatedwith 0 μl/10 ml, 5 μl/10 ml and 10 μl/10 ml Formula I(n). The effect ofFormula I(n) on the migration of Hs578T invasive breast tumor cells wastested using the Membrane Invasion Culture System (MICS) chamber, usingthe protocol described in FIG. 1A. As described in Example 6, FormulaI(n) significantly inhibited Hs578T tumor cell migration.

FIG. 2A is a micrograph of an in vitro scratch assay using fibroblastsshowing the migration of fibroblasts at 0 hours without treatment ofFormula I(n). Confluent cultures of foreskin fibroblasts (Clonetics)were scratched with the tip of a 200 μl pipet tip, rinsed once with PBSand media was changed to serum free media containing 0 μl/10 ml FormulaI(n). Pictures were taken at the indicated time (0, 12, or 24 hours)following scratching and the distance between the migrating lips weremeasured in pixels. FIG. 2B is a micrograph of an in vitro scratch assayusing fibroblasts showing the migration of fibroblasts at 0 hours upontreatment with 5 μl/10 ml Formula I(n). Experiments were conducted asdescribed in FIG. 2A, using 5 μl/10 ml Formula I(n). FIG. 2C is amicrograph of an in vitro scratch assay using fibroblasts showing themigration of fibroblasts at 0 hours upon treatment with 10 μl/10 mlFormula I(n). Experiments were conducted as described in FIG. 2A, using10 μl/10 ml Formula I(n). FIG. 2D is a micrograph of an in vitro scratchassay using fibroblasts showing the migration of fibroblasts at 12 hourswithout treatment of Formula I(n). Experiments were conducted asdescribed in FIG. 2A. FIG. 2E is a micrograph of an in vitro scratchassay using fibroblasts showing the migration of fibroblasts at 12 hoursupon treatment with 5 μl/10 ml Formula I(n). Experiments were conductedas described in FIG. 2A, using 5 μl/10 ml Formula I(n). As described inExample 7, Formula I(n) treated cells showed reduced migration. FIG. 2Fis a micrograph of an in vitro scratch assay using fibroblasts showingthe migration of fibroblasts at 12 hours upon treatment with 10 μl/10 mlFormula I(n). Experiments were conducted as described in FIG. 2A, using10 μl/10 ml Formula I(n). As described in Example 7, Formula I(n)treated cells showed reduced migration. FIG. 2G is a micrograph of an invitro scratch assay using fibroblasts showing the migration offibroblasts at 24 hours without treatment of Formula I(n). Experimentswere conducted as described in FIG. 2A. FIG. 2H is a micrograph of an invitro scratch assay using fibroblasts showing the migration offibroblasts at 24 hours upon treatment with 5 μl/10 ml Formula I(n).Experiments were conducted as described in FIG. 2A, using 5 μl/10 mlFormula I(n). As described in Example 7, Formula I(n) treated cellsshowed reduced migration.

FIG. 3 is a graph of the distance between the migrating lips (asmeasured in pixels) vs. the time (hours) after administration of FormulaI(n). Experiments were performed as described in FIG. 2.

FIG. 4A is a micrograph of preconfluent neonatal foreskin fibroblastcells that have not been treated with Formula I(n). After 24 hours ofmock treatment, cells were stained with phalloidin (actin-green) andpropidium iodide (nuclei-red). FIG. 4B is a micrograph of preconfluentneonatal foreskin fibroblast cells that have been treated with 5 μl/10ml Formula I(n). After 24 hours of treatment, cells were stained withphalloidin (actin-green) and propidium iodide (nuclei-red). FIG. 4C is amicrograph of preconfluent neonatal foreskin fibroblast cells that havebeen treated with 10 μl/10 ml Formula I(n). After 24 hours of treatment,cells were stained with phalloidin (actin-green) and propidium iodide(nuclei-red). FIG. 4D is a micrograph of confluent MDA-MB231 breastcancer cells that have not been treated with Formula I(n). After 24hours of mock treatment, cells were stained with phalloidin(actin-green) and propidium iodide (nuclei-red). FIG. 4E is a micrographof confluent MDA-MB231 breast cancer cells that have been treated with 5μl/10 ml Formula I(n). After 24 hours of treatment, cells were stainedwith phalloidin (actin-green) and propidium iodide (nuclei-red). FIG. 4Fis a micrograph of confluent MDA-MB231 breast cancer cells that havebeen treated with 10 μl/10 ml Formula I(n). After 24 hours of treatment,cells were stained with phalloidin (actin-green) and propidium iodide(nuclei-red). FIG. 4G is a micrograph of preconfluent MDA-MB231 breastcancer cells that have not been treated with Formula I(n). After 24hours of mock treatment, cells were stained with phalloidin(actin-green) and propidium iodide (nuclei-red). FIG. 4H is a micrographof preconfluent MDA-MB231 breast cancer cells that have been treatedwith 5 μl/10 ml Formula I(n). After 24 hours of treatment, cells werestained with phalloidin (actin-green) and propidium iodide (nuclei-red).

FIG. 5 is a western blot showing the inhibition of the phosphorylationof focal adhesion kinase (FAK) in Hs578T cells treated with FormulaI(n). Antibodies that detect the FAK protein that has beenphosphorylated on pTyr576 were used at a dilution of 1:1000 (rabbitanti-FAK[pTyr576] from Biosource Inc., Camarillo, Calif., USA). Asdescribed in Example 9, Formula I(n) induced a concentration dependentdecrease of FAK[Tyr576] phosphorylation.

DETAILED DESCRIPTION OF THE INVENTION

The present invention concerns lipidic furan, pyrrole, and thiophenecompounds, methods, and compositions for treatment of cancer,neurological disorders, and fibrotic disorders.

Definitions

Unless otherwise stated, the following terms used in this application,including the specification and claims, have the definitions givenbelow. As used in the specification and the appended claims, thesingular forms “a,” “an” and “the” include plural referents unless thecontext clearly dictates otherwise. Definition of standard chemistryterms may be found in reference works, including Carey and Sundberg(2007) Advanced Organic Chemistry 5th Ed. Vols. A and B, SpringerScience+Business Media LLC, New York. The practice of the presentinvention will employ, unless otherwise indicated, conventional methodsof synthetic organic chemistry, mass spectroscopy, preparative andanalytical methods of chromatography, protein chemistry, biochemistry,recombinant DNA techniques and pharmacology. Conventional methods oforganic chemistry include those included in March's Advanced OrganicChemistry: Reactions, Mechanisms, and Structure, 6th Edition, M. B.Smith and J. March, John Wiley & Sons, Inc., Hoboken, N.J., 2007.

The term “lipidic furan” as used herein refers a compound comprising afuran ring substituted with at least one branched, straight chain orcyclic hydrocarbon group, preferably a linear hydrocarbon chain, morepreferably comprising a linear hydrocarbon chain comprising one or moreethylenic or acetylenic unsaturations.

As used herein, the term “prodrug” means a compound which whenadministered to a host in vivo is converted into the parent drug. Asused herein, the term “parent drug” means any of the presently describedchemical compounds that are useful to treat any of the disordersdescribed herein, or to control or improve the underlying cause orsymptoms associated with any physiological or pathological disorderdescribed herein in a host, typically a human. Prodrugs can be used toachieve any desired effect, including to enhance properties of theparent drug or to improve the pharmaceutic or pharmacokinetic propertiesof the parent. Prodrug strategies exist which provide choices inmodulating the conditions for in vivo generation of the parent drug, allof which are deemed included herein. Nonlimiting examples of prodrugstrategies include covalent attachment of removable groups, or removableportions of groups, for example, but not limited to acylation,phosphorylation, phosphonylation, phosphoramidate derivatives,amidation, reduction, oxidation, esterification, alkylation, othercarboxy derivatives, sulfoxy or sulfone derivatives, carbonylation oranhydride, among others.

Throughout the specification and claims, a given chemical formula orname shall encompass all optical and stereoisomers, as well as racemicmixtures where such isomers and mixtures exist, unless otherwise noted.

The subject treated is typically a human subject, although it is to beunderstood the methods described herein are effective with respect toother animals, such as mammals and vertebrate species. Moreparticularly, the term subject can include animals used in assays suchas those used in preclinical testing including but not limited to mice,rats, monkeys, dogs, pigs and rabbits; as well as domesticated swine(pigs and hogs), ruminants, equine, poultry, felines, bovines, murines,canines, and the like.

Active Compounds

In one embodiment, the invention is directed to compounds or the use asdescribed herein of such compounds of Formula I, II or III;

wherein;each Z is independently O, S or NR₅;wherein R₁, R₂, R₃ and R₄, are the same or different, separatelyrepresent a hydrogen atom, a C₁-C₃₅ alkyl radical, advantageouslyC₁₀-C₂₂, more advantageously C₁₂-C₂₀, further advantageously C₁₃-C₁₇; aC₁-C₃₅ alkenyl radical, advantageously C₁₀-C₂₂, more advantageouslyC₁₂-C₂₀, further advantageously C₁₃-C₁₇; or a C₁-C₃₅ alkynyl radical,advantageously C₁₀-C₂₂, more advantageously C₁₂-C₂₀, furtheradvantageously C₁₃-C₁₇, and wherein the alkyl, alkenyl or alkynyl moietyis optionally substituted with one or more halogens (F, Cl, Br, or I,and more typically F) and/or by one or more moieties selected from thegroup consisting of epoxide (e.g, an oxygen divalently linked to thecarbon chain), hydroxyl or protected hydroxyl (OR₅), thiol or protectedthiol (SR₅), amine (NR₅R₆), aldehyde (—CHO), ketone (—COR₅), acetyl(—O—CO—R₅), or ester (—C(O)OR₅) function and wherein R₅ and R₆separately represent a hydrogen atom, a C₁-C₃₅, more typically a C₁ toC₂ alkyl radical, advantageously C₁₀-C₂₂, more advantageously C₁₂-C₂₀,further advantageously C₁₃-C₁₇, or a C₁-C₃₅ alkenyl radical,advantageously C₁₀-C₂₂, more advantageously C₁₂-C₂₀, or furtheradvantageously C₁₃-C₁₇.

In some embodiments, the compound has Formula I(a):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄=C₁₃-C₁₉ alkyl optionally substituted as defined above.

In some embodiments, the compound has Formula I(b):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a straight C₉-C₂₀ unsaturated alkyl chain with a single doublebond of the formula —CH═CH(CH₂)_(m)CH₃;optionally substituted as defined above;wherein;m=6 to 17.

In some embodiments, the compound has Formula I(c):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a straight C₁₃-C₁₉ unsaturated alkyl chain with a single doublebond of the formula —CH═CH(CH₂)_(m)CH₃;optionally substituted as defined above;wherein;m=10 to 16.

In some embodiments, the compound has Formula I(d):

wherein;

Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a straight C₉-C₂₀ unsaturated alkyl chain with a single doublebond of the formula —(CH₂)_(n)—CH═CH(CH₂)_(m)CH₃;optionally substituted as defined above;whereinn=1 to 17;m=1 to 17.

In some embodiments, the compound has Formula I(e):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a straight C₁₃-C₁₉ unsaturated alkyl chain with a single doublebond of the formula —(CH₂)_(n)—CH═CH(CH₂)_(m)CH₃;optionally substituted as defined above;whereinn=1 to 15;m=1 to 15.

In some embodiments, the compound has Formula I(f):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a straight C₉-C₂₀ unsaturated alkyl chain with a single doublebond of the formula —(CH₂)_(m)CH═CH₂;optionally substituted as defined above;wherein;m=7 to 18.

In some embodiments, the compound has Formula I(g):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a straight C₁₃-C₁₉ unsaturated alkyl chain with a single doublebond of the formula —(CH₂)_(m)CH═CH₂;optionally substituted as defined above;wherein;m=11 to 17.

In some embodiments, the compound has Formula I(h):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a C₉-C₂₀ alkyl chain comprising two or more double bondsoptionally substituted as defined above.

In some embodiments, the compound has Formula I(i):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a C₁₃-C₁₉ alkyl chain comprising two or more double bondsoptionally substituted as defined above.

In some embodiments, the compound has Formula I(j):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a C₉-C₂₀ alkyl chain comprising one or more triple bondsoptionally substituted as defined above.

In some embodiments, the compound has Formula I(k):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a C₁₃-C₁₉ alkyl chain comprising one or more triple bondsoptionally substituted as defined above.

In some embodiments, the compound has Formula I(l):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a C₉-C₂₀ alkyl chain comprising one or more double bonds and oneor more triple bonds optionally substituted as defined above.

In some embodiments, the compound has Formula I(m):

wherein;Z is independently O, S, NR₅;R₁=R₂=R₃=H;R₄ is a C₁₃-C₁₉ alkyl chain comprising one or more double bonds and oneor more triple bonds optionally substituted as defined above.

In an alternative embodiment, the furan, thiophene or pyrrole can befused to another heterocyclic or heteroaromatic moiety to produce amulti-ring core, for example, benzofuran, benzothiophene, or indole,which may optionally be substituted with one or more functional groups,preferably with one or more alkyl, alkoxy, halo, or hydroxysubstituents.

In one embodiment, the compound has Formula I(n):

In one embodiment, the compound has Formula I(o):

In one embodiment, the compound has Formula I(p):

In one embodiment, the compound has Formula I(q):

In one embodiment, the compound has Formula I(r):

In one embodiment, the compound has Formula I(s):

In one embodiment, the compound has Formula I(t):

In one embodiment, the compound has Formula I(u):

In one embodiment, the compound has Formula I(v):

In one embodiment, the compound has Formula I(w):

In one embodiment, the compound has Formula I(x):

In one embodiment, the compound has Formula I(y):

In one embodiment, the compound has Formula I(z):

In one embodiment, the compound has the Formula I(aa):

In one embodiment, the compound has the Formula I(ab):

In one embodiment, the compound has the Formula I(ac):

In one embodiment, the compound has the Formula I(ad):

In one embodiment, the compound has the Formula I(ae):

In one embodiment, the compound has the Formula I(af):

In one embodiment, the compound has the Formula I(ag):

In one embodiment, the compound has the Formula I(ah):

In one embodiment, the compound has the Formula I(ai):

In one embodiment, the compound has the Formula I(aj):

In one embodiment, the compound has the Formula I(ak):

In one embodiment, the compound has the Formula I(al):

In one embodiment, the compound has the Formula I(am):

In one embodiment, the compound has the Formula I(an):

In one embodiment, the compound has the Formula I(ao):

In one embodiment the compound of Formula I is 2-(nonanyl)furan.

In one embodiment the compound of Formula I is 2-(dodecyl)furan.

In one embodiment the compound of Formula I is 2-(tridecyl)furan.

In one embodiment the compound of Formula I is 2-(tetradecyl)furan.

In one embodiment the compound of Formula I is 2-(pentadecyl)furan,

In one embodiment the compound of Formula I is 2-(hexadecyl)furan,

In one embodiment the compound of Formula I is 2-(heptadecyl)furan

In one embodiment the compound of Formula I is 2-(octadecyl)furan).

In one embodiment the compound of Formula I is 2-(nonadecyl)furan).

In one embodiment the compound of Formula I is 2-(8Z-pentadecenyl)furan.

In one embodiment the compound of Formula I is 2-(1E-pentadecenyl)furan.

In one embodiment the compound of Formula I is 2-(1Z-pentadecenyl)furan.

In one embodiment the compound of Formula I is 2-(12-tridecenyl)furan.

In one embodiment the compound of Formula I is2-(8Z,11Z-heptadecadienyl)furan.

In one embodiment the compound of Formula I is2-(8Z,11Z-heptadecadienyl)thiophene.

In one embodiment the compound of Formula I is2-(8Z,11Z-heptadecadienyl)pyrrole.

In one embodiment the compound of Formula I is lignoceric furan.

In one embodiment the compound of Formula I is lauroleic furan.

In one embodiment the compound of Formula I is palmitoleic furan.

In one embodiment the compound of Formula I is cis-vaccenic furan.

In one embodiment the compound of Formula I is erucic furan.

In one embodiment the compound of Formula I is nervonic furan.

In one embodiment the compound of Formula I is arachidonic furan.

In one embodiment the compound of Formula I is crepenynic furan.

Certain furan, pyrrolo, and thiophene derivatives have been of interestin the petroleum, electronic and pharmaceutical industries for certaindisorders. Notwithstanding this research, it has now been surprisinglydiscovered that the presently disclosed active compounds can beadvantageously used to treat solid tumors. Thus this invention can beused to treat cancers including but not limited to solid tumors such asbreast, colon, lung, bladder, kidney, liver or pancreatic cancer. It hasalso not been known to use the presently disclosed active compounds totreat serious neurological disorders such as Alzheimer's, Parkinson's,or dementia, nor fibrotic disorders.

In regard to furan derivatives, Kashman et al. first reported theavocadofurans as a new class of phytochemicals. See, Kashman, Y, et al.,“New Compounds from Avocado Pear”, Tetrahedron, 25:4617-4631 (1969) andKashman, Y, et al., “Six New C17-olefinic and Acetylenic OxygenatedCompounds from Avocado Pear”, Isr. J. Chem., 7:173-176. The authorsisolated 2-(trideca-12-ynyl)furan and 2-(trideca-12-enyl)furan from P.americana fruit and seeds. Magalhaes et al. subsequently identifiedseveral other 2-alkylfurans with C₁₃ mono- and diunsaturated side chainsfrom methanol extracts of avocado seeds [Persea gratissima Gartn. (syn.P. americana)] from Brazil. See, Magalhaes et al., “The avocatins—a newclass of natural products”, An. Acd. Bras. Cienc. 42(suppl):45-48(1970).

Neeman et al. tested a group of eight long-chain aliphatic compoundsfrom avocados for activity against 13 species of bacteria and a yeastand reported that 2-(trideca-12-enyl)furan inhibited growth of Bacillussubtilis (Ehrenberg) Cohn and Staphylococcus aureus Rosenbach. See,Neeman, I, et al., “New antibacterial agent isolated from the avocadopear”, Appl. Microbiol., 19:470-473, (1970). Murakoshi et al. tested2-(8Z, 11Z-heptadecadienyl)furan produced by acid-catalyzed dehydrationof persin from avocado leaves against silkworm larvae, B. mori, andfound no activity at concentrations in the diet up to 300 μg/g. See,Murakoshi et al., J. Appl. Entomol. Zool. 20:87-91 (1976).Rodriguez-Saona, C., et al., disclosed that avocadofurans are active asantifeedants. See, Rodriguez-Saona, C., et al., “Novel Antifeedant andInsecticidal Compounds from Avocado Idioblast Cell Oil”, J. Chem. Ecol.,24:867-889 (1998). In addition, the authors disclosed the synthesis ofC₁₅ and C₁₇ 2-(alkyl)furans. The compounds were synthesized by couplingthe appropriate bromoalkane with 2-lithiofuran in THF.2-(1E-pentadecenyl)furan and its Z isomer were prepared by Wittigreaction of the ylide prepared from tetradecyl triphenylphosphoniumbromide with furfural to accord a 7:3 mixture of Z and E isomers.

The compound 2-(8Z,11Z-heptadecadienyl)furan was synthesized as follows.Linoleic acid was converted to the tetrabromide using bromine in diethylether. The carboxylic acid was next decarboxylated and converted to analkyl bromide using a modified Hunsdiecker reaction. Regeneration of thediene moiety with zinc powder in THE yielded a bromo diene that wasconverted to the corresponding alkyl iodide and subsequently coupledwith 2-lithiofuran to afford the doubly unsaturated avacadofuran.

U.S. Pat. No. 5,468,490 to Huber, S. R. and Counts, D. F. discloses thelipidic furan, 17-(2-furanyl-8-11-cis-cis-heptadecadiene, also referredto as 2-(8Z,11Z-heptadecadienyl)furan), with beneficial effects on theepidermis and dermis of the skin. U.S. Pat. No. 5,514,709 to Counts, D.F. and Huber, R. discloses lipid furans, 2-alkyl furans, and theirspecificity to types I and III collagen, both of which are present inlarge amounts in the skin and mucosal tissues. Rodriguez-Saona, C. etal. discloses the synthesis of 2-(pentadecyl)furan and2-(heptadecyl)furan. See, Rodriquez-Saona, C., et al., “Avocadofuransand Their Tetrahydrofuran Analogues: Comparison of Growth Inhibitory andInsecticidal Activity”, J. Agric. Food Chem., 48:3642-3645 (2000). U.S.Pat. Application No. 2004/0018258 to Piccirilli, A. and Legrand, J.discloses the process for obtaining a furan lipid-rich unsaponifiablematerial from avocado. U.S. Pat. Application No. 2008/0219937 to Msika,P. and Piccardi, N. discloses the use of a cosmetic composition withdepigmenting or lightening action comprising as active at least one2-alkyl furan. U.S. Pat. Application No. 2008/0081837 to Piccirilli, A.,et al., disclose a method for preventing and/or treating diabetes using2-alkyl furans wherein the 2-position is substituted with a C₁-C₃₅alkyl, C₁-C₃₅ alkenyl, or C₁-C₃₅ alkynyl substituent. U.S. Pat. No.7,589,121 to Piccirilli, A. et al., discloses the use of alkyl furansfor the treatment of obesity and obesity. U.S. Pat. Application No.2013/0183289 to Gorelik, L. et al discloses the use avocadanofuran forthe treatment of a DNA virus.

In regard to pyrroles, U.S. Pat. No. 5,082,856 to Taniguchi, M. et aldiscloses pyrrolecarboxylic acid derivatives for the treatment ofhyperlipemia and arteriosclerosis. The authors disclose the synthesis of2,5-disubstituted, and 2,4-disubstituted pyrroles. For example, inMethod 1 of the '856 patent, pyrrole is reacted with a methyl or ethylmagnesium halide and the magnesium salt is reacted with an acyl chlorideto afford a 2-acylpyrrole. The ketone is subjected to Wolff-Kishnerreduction conditions to afford the 2-alkylpyrrole. The 2-alkylpyrrole isthen treated with a Grignard reagent followed by ethyl chlorocarbonateto afford 5-alkylpyrrole-2-carboxylate. The ester can be converted tothe corresponding carboxylic acid by hydrolyzing the ester understandard conditions. In Method 2 of the '856 patent, ethylpyrrole-3-carboxylate is reacted with an acyl chloride in the presenceof a Lewis acid to obtain the 5-acyl-pyrrole-3-carboxylate. Theresulting ketone is then subjected to Wolff-Kishner reduction conditionsto afford an ethyl 5-alkylpyrrole-3-carboxylate. The ester can then beconverted to the corresponding carboxylic acid by hydrolyzing the esterunder standard conditions.

Mycalazol and mycalazal analogs having antiproliferating activity beendisclosed by Mohamed, Y. M. A. and Hansen, T. V. See, Mohamed, Y. M. A.and Hansen, T. V., “Synthesis of mycalazol and mycalazal analogs withpotent antiproliferating activities”, Pure Appl. Chem., 83(3):489-493(2011). Using pyrrole 2-carboxaldehyde as the starting material, theauthors treated the aldehyde with dimethylamine to form an azafulvene inquantitative yield. The compound was treated with n-butyllithium andexcess n-tributyltin chloride to produce5-(tri-n-butylstannyl)pyrrole-2-carboxaldehyde. Acid chlorides underwentStille cross-coupling reactions with the stannyl pyrrole to afford2,5-disubstituted pyrroles.

Papireddy, K. et al. have recently disclosed the antimalarial activityof prodiginines. In this paper, the authors disclosed the synthesis of2-alkylated pyrroles and 3-alkylated pyrroles. The intermediates weresubsequently used to generate synthetic analogs of prodiginines. Inorder to generate 2-alkylated pyrroles, pyrrole was acylated using zincpowder to afford 2-acylpyrroles. The 2-acylpyrroles were reduced to thecorresponding 2-alkylated pyrroles using an excess of sodium borohydridein 2-propanol under reflux. In order to generate 3-alkylated pyrroles,pyrrole was treated with phenylsulfonyl chloride in the presence ofsodium hydroxide to provide N-phenylsulfonylpyrrole. Use of aluminumtrichloride permitted the regioselective acylation of theN-phenylsulfonylpyrrole at the 3 position with an acyl chloride toafford N-phenylsulfonyl-3-acylpyrroles. TheN-phenylsulfonyl-3-acylpyrroles were hydrolyzed under basic conditionsto afford 3-acylpyrroles. In the final step, 3-acylpyrroles were reducedto afford 3-alkylpyrroles using sodium borohydride under reflux. See,Papireddy, K. et al., “Antimalarial Activity of Natural and SyntheticProdiginines”, J. Med. Chem., 54:5296-5306 (2011).

In regard to thiophene derivatives, 2,5-disubstituted thiophene analogswere disclosed by Buu-Hoi, N. P, et al. as potential tuberculostaticcompounds. See, Buu-Hoi, N. P, et al.,

Isotopic Substitution

The present invention includes compounds and the use of compounds withdesired isotopic substitutions of atoms, at amounts above the naturalabundance of the isotope, i.e., enriched. Isotopes are atoms having thesame atomic number but different mass numbers, i.e., the same number ofprotons but a different number of neutrons. By way of general exampleand without limitation, isotopes of hydrogen, for example, deuterium(²H) and tritium (³H) may be used anywhere in described structures.Alternatively or in addition, isotopes of carbon, e.g., ¹³C and ¹⁴C, maybe used. A preferred isotopic substitution is deuterium for hydrogen atone or more locations on the molecule to improve the performance of thedrug. The deuterium can be bound in a location of bond breakage duringmetabolism (an α-deuterium kinetic isotope effect) or next to or nearthe site of bond breakage (a β-deuterium kinetic isotope effect).

Substitution with isotopes such as deuterium can afford certaintherapeutic advantages resulting from greater metabolic stability, suchas, for example, increased in vivo half-life or reduced dosagerequirements. Substitution of deuterium for hydrogen at a site ofmetabolic break down can reduce the rate of or eliminate the metabolismat that bond. At any position of the compound that a hydrogen atom maybe present, the hydrogen atom can be any isotope of hydrogen, includingprotium (H), deuterium (²H) and tritium (3H). Thus, reference herein toa compound encompasses all potential isotopic forms unless the contextclearly dictates otherwise.

The term “isotopically-labeled” analog refers to an analog that is a“deuterated analog”, a “¹³C-labeled analog,” or a“deuterated/¹³C-labeled analog.” The term “deuterated analog” means acompound described herein, whereby a H-isotope, i.e., hydrogen/protium(H), is substituted by a H-isotope, i.e., deuterium (²H). Deuteriumsubstitution can be partial or complete. Partial deuterium substitutionmeans that at least one hydrogen is substituted by at least onedeuterium. In certain embodiments, the isotope is 90, 95 or 99% or moreenriched in an isotope at any location of interest. In some embodimentsit is deuterium that is 90, 95 or 99% enriched at a desired location.

Therapeutic Treatment of Cancers

Compounds, methods, and compositions are provided to treat a cancer.Improved compounds, methods, and compositions are provided forinhibiting cellular migration of cancer cells. Improved compounds,methods, and compositions are provided that inhibit invasive cancers.Improved compounds, methods, and compositions are provided forinhibiting abnormal cellular proliferation. In one embodiment, thecancer is a solid tumor. In one embodiment, the solid tumor is anon-skin tumor.

In one embodiment of the invention, a compound for the treatment ofcancer is selected from the compounds of Formula I, II or III asdescribed herein, or a pharmaceutically acceptable composition, salt,isotopic analog, or prodrug thereof. In one non-limiting example, thecancer is treated with Formula I(a), Formula I(b), Formula I(c), FormulaI(d), Formula I(e), Formula I(f), Formula I(g), Formula I(h), FormulaI(i), Formula I(j), Formula I(k), Formula I(l), Formula I(m), FormulaI(n), Formula I(o), Formula I(p), Formula I(q), Formula I(r), FormulaI(s), Formula I(t), Formula I(u), Formula I(v), Formula I(w), FormulaI(x), Formula I(y), Formula I(z), Formula I(aa), Formula I(ab), FormulaI(ac), Formula I(ad), Formula I(ae), Formula I(af), Formula I(ag),Formula I(ah), Formula I(ai), Formula I(aj), Formula I(ak), FormulaI(al), Formula I(am), Formula I(an), or Formula I(ao) as describedherein, or a pharmaceutically acceptable composition, salt, isotopicanalog, or prodrug thereof.

According to this invention, the active ingredient is not provided as abotanical extract mixture or combination, but instead the activecompound is delivered in a highly pure form. In one embodiment, theinvention is a dosage form for the treatment of a solid tumor, which isa non-skin derived tumor or disorder, wherein the active compound has apurity of at least 96%, 97%, 98%, or 99%, without respect to fillers,stabilizers, or other inert or inactive ingredients. In an alternativeembodiment, the dosage form has two or more active ingredients, whereinonly one of the active ingredients is selected from compounds of FormulaI, II or III as described herein. In a further alternative embodiment,the dosage form has two or more active ingredients selected fromcompounds of Formula I, II or III as described herein, wherein eachactive compound has a purity of at least 96%, 97%, 98%, or 99%, withoutrespect to fillers, stabilizers, or other inert or inactive ingredients.

Further provided herein is a method of reducing growth of a solid tumorin a subject, comprising administering the active compounds describedherein. A tumor can be a primary tumor or a metastatic tumor. In oneaspect, a solid tumor is, for example, lung cancer (including lungadenocarcinoma, basal cell carcinoma, squamous cell carcinoma, largecell carcinoma, bronchioloalveolar carcinoma, bronchiogenic carcinoma,non-small-cell carcinoma, small cell carcinoma, mesothelioma); breastcancer (including ductal carcinoma, lobular carcinoma, inflammatorybreast cancer, clear cell carcinoma, mucinous carcinoma, serosalcavities breast carcinoma); colorectal cancer (colon cancer, rectalcancer, colorectal adenocarcinoma); anal cancer; pancreatic cancer(including pancreatic adenocarcinoma, islet cell carcinoma,neuroendocrine tumors); prostate cancer; prostate adenocarcinoma;ovarian carcinoma (ovarian epithelial carcinoma or surfaceepithelial-stromal tumor including serous tumor, endometrioid tumor andmucinous cystadenocarcinoma, sex-cord-stromal tumor); liver and bileduct carcinoma (including hepatocellular carcinoma, cholangiocarcinoma,hemangioma); esophageal carcinoma (including esophageal adenocarcinomaand squamous cell carcinoma); oral and oropharyngeal squamous cellcarcinoma; salivary gland adenoid cystic carcinoma; bladder cancer;bladder carcinoma; carcinoma of the uterus (including endometrialadenocarcinoma, uterine papillary serous carcinoma, uterine clear-cellcarcinoma, uterine sarcomas and leiomyosarcomas, mixed mulleriantumors); glioma, glioblastoma, medullablastoma, and other tumors of thebrain; kidney cancers (including renal cell carcinoma, clear cellcarcinoma, Wilm's tumor); cancer of the head and neck (includingsquamous cell carcinomas); cancer of the stomach (gastric cancers,stomach adenocarcinoma, gastrointestinal stromal tumor); testicularcancer; germ cell tumor; neuroendocrine tumor; cervical cancer;carcinoids of the gastrointestinal tract, breast, and other organs;signet ring cell carcinoma; mesenchymal tumors including sarcomas,fibrosarcomas, haemangioma, angiomatosis, haemangiopericytoma,pseudoangiomatous stromal hyperplasia, myofibroblastoma, fibromatosis,inflammatory myofibroblastic tumor, lipoma, angiolipoma, granular celltumor, neurofibroma, schwannoma, angiosarcoma, liposarcoma,rhabdomyosarcoma, osteosarcoma, leiomyoma, leiomysarcoma, or melanoma.In one embodiment, a solid tumor is, for example, a colon tumor, anovarian tumor, a lung tumor, an esophageal tumor, a breast tumor, aprostate tumor, a carcinoma. In one embodiment of the invention, thecancer is treated with a compound selected from the compounds of FormulaI, II or III as described herein, or a pharmaceutically acceptablecomposition, salt, isotopic analog, or prodrug thereof. In onenon-limiting example, the cancer is treated with Formula I(a), FormulaI(b), Formula I(c), Formula I(d), Formula I(e), Formula I(f), FormulaI(g), Formula I(h), Formula I(i), Formula I(j), Formula I(k), FormulaI(l), Formula I(m), Formula I(n), Formula I(o), Formula I(p), FormulaI(q), Formula I(r), Formula I(s), Formula I(t), Formula I(u), FormulaI(v), Formula I(w), Formula I(x), Formula I(y), Formula I(z), FormulaI(aa), Formula I(ab), Formula I(ac), Formula I(ad), Formula I(ae),Formula I(af), Formula I(ag), Formula I(ah), Formula I(ai), FormulaI(aj), Formula I(ak), Formula I(al), Formula I(am), Formula I(an), orFormula I(ao) as described herein, or a pharmaceutically acceptablecomposition, salt, isotopic analog, or prodrug thereof.

In one embodiment, the cancer is an invasive breast cancer. Invasivebreast cancer has spread from the original site (either the milk ductsor lobules) into the surrounding breast tissue, and possibly spread tothe lymph nodes and/or other parts of the body. Thus, invasive breastcancers have a poorer prognosis than ductal carcinoma in situ (DCIS),which is a non-invasive breast cancer. Using a microscope to look at thetissue removed during a biopsy, a pathologist can determine whether atumor is DCIS or invasive breast cancer through analysis of biopsies.The most common type is invasive ductal carcinoma (also calledinfiltrating ductal carcinoma and less commonly, invasive carcinoma ofno special type or invasive carcinoma not otherwise specified). Invasiveductal carcinoma accounts for 50 to 75 percent of all breast cancers.Invasive lobular carcinoma is the next most common type and accounts forabout 10 to 15 percent of cases. Tubular carcinoma and mucinous(colloid) carcinoma are less common types of invasive breast cancer thattend to have a good prognosis. In one embodiment of the invention, thebreast cancer is treated with a compound selected from the compounds ofFormula I, II or III as described herein, or a pharmaceuticallyacceptable composition, salt, isotopic analog, or prodrug thereof. Inone non-limiting example, the breast cancer is treated with FormulaI(a), Formula I(b), Formula I(c), Formula I(d), Formula I(e), FormulaI(f), Formula I(g), Formula I(h), Formula I(i), Formula I(j), FormulaI(k), Formula I(l), Formula I(m), Formula I(n), Formula I(o), FormulaI(p), Formula I(q), Formula I(r), Formula I(s), Formula I(t), FormulaI(u), Formula I(v), Formula I(w), Formula I(x), Formula I(y), FormulaI(z), Formula I(aa), Formula I(ab), Formula I(ac), Formula I(ad),Formula I(ae), Formula I(af), Formula I(ag), Formula I(ah), FormulaI(ai), Formula I(aj), Formula I(ak), Formula I(al), Formula I(am),Formula I(an), or Formula I(ao) as described herein, or apharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof.

In one embodiment provided herein is a method of treating a skin cancercomprising administering the active compounds described herein. In oneembodiment, the skin cancer is selected from the group consisting of allnon-melanoma skin cancers such as squamous cell carcinoma and basal cellcarcinoma. In one embodiment, the skin cancer is a melanoma. In oneembodiment of the invention, the skin cancer is treated with a compoundselected from the compounds of Formula I, II or III as described herein,or a pharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof. In one non-limiting example, the skin cancer is treatedwith Formula I(a), Formula I(b), Formula I(c), Formula I(d), FormulaI(e), Formula I(f), Formula I(g), Formula I(h), Formula I(i), FormulaI(j), Formula I(k), Formula I(l), Formula I(m), Formula I(n), FormulaI(o), Formula I(p), Formula I(q), Formula I(r), Formula I(s), FormulaI(t), Formula I(u), Formula I(v), Formula I(w), Formula I(x), FormulaI(y), Formula I(z), Formula I(aa), Formula I(ab), Formula I(ac), FormulaI(ad), Formula I(ae), Formula I(af), Formula I(ag), Formula I(ah),Formula I(ai), Formula I(aj), Formula I(ak), Formula I(al), FormulaI(am), Formula I(an), or Formula I(ao) as described herein, or apharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof.

In certain aspects, compounds, methods, and composition are provided foruse as chemotherapeutics for treating and/or preventing cancer cell orpre-cancerous lesion expansion, proliferation, o activation, in apatient in need thereof, comprising administering a pharmaceuticallyeffective amount of the compounds described herein. In certainembodiments, compounds, methods, and composition are provided for use aschemotherapeutics for treating and/or preventing cancer cell metastasisor invasion in a patient in need thereof, comprising administering apharmaceutically effective amount of the compounds described herein. Incertain embodiments, compounds, methods, and composition are providedfor use as chemotherapeutics for treating and/or preventing cancer cellmigration in a patient in need thereof, comprising administering apharmaceutically effective amount of the compounds described herein. Incertain embodiments, compounds, methods, and composition are providedfor use as chemotherapeutics for treating and/or preventing cytoskeletalreorganization of the cancer cell in a patient in need thereof,comprising administering a pharmaceutically effective amount of thecompounds described herein.

In one embodiment of the invention, a compound(s) of Formula I, having apurity of greater than 98%, is incorporated with an excipient(s) fortopical application as a medicament to treat cancerous lesions of theskin. In one embodiment, the amount of the compound of Formula I fortopical application ranges from about 0.1% to about 100%. In oneembodiment, the amount of the compound of Formula I for topicalapplication ranges from about 1% to about 10%. In another embodiment,the amount of the compound of Formula I for topical application rangesfrom about 1% to about 5%. In one embodiment of the invention, theexcipient is an oil of natural or synthetic origin. In one embodiment,the cancers of the skin include, but are not limited to, non-melanomaskin cancers, squamous cell carcinoma and basal cell carcinoma.

Therapeutic Treatment of Pre-Cancerous Lesions

Actinic keratoses (AKs) are common skin lesions heralding an increasedrisk of developing squamous cell carcinoma (SCC) and other skinmalignancies, arising principally due to excessive ultraviolet (UV)exposure. They are predominantly found in fair-skinned individuals, andincreasingly, are a problem of the immunosuppressed. Actinic keratoses(AKs) may regress spontaneously, remain stable or transform to invasiveSCC.

In one embodiment provided herein is a method of treating and/orpreventing a pre-cancerous lesion of the skin, comprising administeringthe active compounds described herein. In one embodiment, thepre-cancerous lesion of the skin is selected from the group consistingof all pre-cancerous lesions, such as actinic keratoses and leukoplakia.In one embodiment of the invention, a compound for the treatment of apre-cancerous lesion of the skin is selected from the compounds ofFormula I, II or III as described herein, or a pharmaceuticallyacceptable composition, salt, isotopic analog, or prodrug thereof. Inone non-limiting example, the pre-cancerous lesion of the skin istreated with Formula I(a), Formula I(b), Formula I(c), Formula I(d),Formula I(e), Formula I(f), Formula I(g), Formula I(h), Formula I(i),Formula I(j), Formula I(k), Formula I(l), Formula I(m), Formula I(n),Formula I(o), Formula I(p), Formula I(q), Formula I(r), Formula I(s),Formula I(t), Formula I(u), Formula I(v), Formula I(w), Formula I(x),Formula I(y), Formula I(z), Formula I(aa), Formula I(ab), Formula I(ac),Formula I(ad), Formula I(ae), Formula I(af), Formula I(ag), FormulaI(ah), Formula I(ai), Formula I(aj), Formula I(ak), Formula I(al),Formula I(am), Formula I(an), or Formula I(ao) as described herein, or apharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof.

According to this invention, the active ingredient is not provided as abotanical extract mixture or combination, but instead the activecompound is delivered in a highly pure form. In one embodiment, theinvention is a dosage form for the treatment of a precancerous lesion,wherein the active compound has a purity of at least 96%, 97%, 98%, or99%, without respect to fillers, stabilizers, or other inert or inactiveingredients. In an alternative embodiment, the dosage form has two ormore active ingredients, wherein only one of the active ingredients isselected from compounds of Formula I, II or III as described herein. Ina further alternative embodiment, the dosage form has two or more activeingredients selected from compounds of Formula I, II or III as describedherein, wherein each active compound has a purity of at least 96%, 97%,98%, or 99%, without respect to fillers, stabilizers, or other inert orinactive ingredients.

In one embodiment of the invention, a compound(s) of Formula I, having apurity of greater than 98%, is incorporated with an excipient(s) fortopical application as a medicament to treat pre-cancerous lesions ofthe skin. In one embodiment, the amount of the compound of Formula I fortopical application ranges from about 0.1% to about 100%. In oneembodiment, the amount of the compound of Formula I for topicalapplication ranges from about 1% to about 10%. In another embodiment,the amount of the compound of Formula I for topical application rangesfrom about 1% to about 5%. In one embodiment of the invention, theexcipient is an oil of natural or synthetic origin. In one embodimentthe pre-cancerous lesions of the skin include, but are not limited to,actinic keratosis and leukoplakia. In one embodiment, the pre-cancerouslesion is leukoplakia.

Cancer Combination Therapy

In one aspect of the invention, the compounds disclosed herein can bebeneficially administered in combination with another therapeuticregimen for beneficial, additive, or synergistic effect.

In one embodiment, a compound/method of the present invention is used incombination with another therapy to treat cancer. In some embodiments,the compound can be administered to the subject in combination withother chemotherapeutic agents. If convenient, the compounds describedherein can be administered at the same time as another chemotherapeuticagent, in order to simplify the treatment regimen. In some embodiments,the compound and the other chemotherapeutic can be provided in a singleformulation. In one embodiment, the use of the compounds describedherein is combined in a therapeutic regime with other agents. Suchagents may include, but are not limited to, tamoxifen, midazolam,letrozole, bortezomib, anastrozole, goserelin, an mTOR inhibitor, a PI3kinase inhibitors, dual mTOR-PI3K inhibitors, MEK inhibitors, RASinhibitors, ALK inhibitors, HSP inhibitors (for example, HSP70 and HSP90 inhibitors, or a combination thereof), BCL-2 inhibitors, apopototicinducing compounds, AKT inhibitors, including but not limited to,MK-2206, GSK690693, Perifosine, (KRX-0401), GDC-0068, Triciribine,AZD5363, Honokiol, PF-04691502, and Miltefosine, PD-1 inhibitorsincluding but not limited to, Nivolumab, CT-011, MK-3475, BMS936558, andAMP-514 or FLT-3 inhibitors, including but not limited to, P406,Dovitinib, Quizartinib (AC220), Amuvatinib (MP-470), Tandutinib(MLN518), ENMD-2076, and KW-2449, or combinations thereof. Examples ofmTOR inhibitors include but are not limited to rapamycin and itsanalogs, everolimus (Afinitor), temsirolimus, ridaforolimus, sirolimus,and deforolimus. Examples of P13 kinase inhibitors include but are notlimited to Wortmannin, demethoxyviridin, perifosine, idelalisib, PX-866,IPI-145, BAY 80-6946, BEZ235, RP6503, TGR 1202 (RP5264), MLN1117(INK1117), Pictilisib, Buparlisib, SAR245408 (XL147), SAR245409 (XL765),Palomid 529, ZSTK474, PWT33597, RP6530, CUDC-907, and AEZS-136. Examplesof MEK inhibitors include but are not limited to Tametinib, Selumetinib,MEK162, GDC-0973 (XL518), and PD0325901. Examples of RAS inhibitorsinclude but are not limited to Reolysin and siG12D LODER. Examples ofALK inhibitors include but are not limited to Crizotinib, AP26113, andLDK378. HSP inhibitors include but are not limited to Geldanamycin or17-N-Allylamino-17-demethoxygeldanamycin (17AAG), and Radicicol.

In one embodiment, a compound described herein can be combined with achemotherapeutic selected from, but are not limited to, Imatinibmesylate (Gleevac®), Dasatinib (Sprycel®), Nilotinib (Tasigna®),Bosutinib (Bosulif®), Trastuzumab (Herceptin®), Pertuzumab (Perjeta™),Lapatinib (Tykerb®), Gefitinib (Iressa®), Erlotinib (Tarceva®),Cetuximab (Erbitux®), Panitumumab (Vectibix®), Vandetanib (Caprelsa®),Vemurafenib (Zelboraf®), Vorinostat (Zolinza®), Romidepsin (Istodax®),Bexarotene (Tagretin®), Alitretinoin (Panretin®), Tretinoin (Vesanoid®),Carfilizomib (Kyprolis™), Pralatrexate (Folotyn®), Bevacizumab(Avastin®), Ziv-aflibercept (Zaltrap®), Sorafenib (Nexavar®), Sunitinib(Sutent®), Pazopanib (Votrient®), Regorafenib (Stivarga®), andCabozantinib (Cometriq™), Vincristine (Oncovin®) or liposomalvincristine (Marqibo®), Daunorubicin (daunomycin or Cerubidine®) ordoxorubicin (Adriamycin®), Cytarabine (cytosine arabinoside, ara-C, orCytosar®), L-asparaginase (Elspar®) or PEG-L-asparaginase (pegaspargaseor Oncaspar®), Etoposide (VP-16), Teniposide (Vumon®), 6-mercaptopurine(6-MP or Purinethol®), Methotrexate, Cyclophosphamide (Cytoxan®),Prednisone, Dexamethasone (Decadron), imatinib (Gleevec®), dasatinib(Sprycel®), nilotinib (Tasigna®), bosutinib (Bosulif®), and ponatinib(Iclusig™). Examples of additional suitable chemotherapeutic agentsinclude but are not limited to 1-dehydrotestosterone, 5-fluorouracildecarbazine, 6-mercaptopurine, 6-thioguanine, actinomycin D, adriamycin,aldesleukin, alkylating agents, allopurinol sodium, altretamine,amifostine, anastrozole, anthramycin (AMC)), anti-mitotic agents,cis-dichlorodiamine platinum (II) (DDP) cisplatin), diamino dichloroplatinum, anthracyclines, antibiotics, antimetabolites, asparaginase,BCG live (intravesical), betamethasone sodium phosphate andbetamethasone acetate, bicalutamide, bleomycin sulfate, busulfan,calcium leucouorin, calicheamicin, capecitabine, carboplatin, lomustine(CCNU), carmustine (BSNU), Chlorambucil, Cisplatin, Cladribine,Colchicin, conjugated estrogens, Cyclophosphamide, Cyclothosphamide,Cytarabine, Cytarabine, cytochalasin B, Cytoxan, Dacarbazine,Dactinomycin, dactinomycin (formerly actinomycin), daunirubicin HCL,daunorucbicin citrate, denileukin diftitox, Dexrazoxane,Dibromomannitol, dihydroxy anthracin dione, Docetaxel, dolasetronmesylate, doxorubicin HCL, dronabinol, E. coli L-asparaginase, emetine,epoetin-α, Erwinia L-asparaginase, esterified estrogens, estradiol,estramustine phosphate sodium, ethidium bromide, ethinyl estradiol,etidronate, etoposide citrororum factor, etoposide phosphate,filgrastim, floxuridine, fluconazole, fludarabine phosphate,fluorouracil, flutamide, folinic acid, gemcitabine HCL, glucocorticoids,goserelin acetate, gramicidin D, granisetron HCL, hydroxyurea,idarubicin HCL, ifosfamide, interferon α-2b, irinotecan HCL, letrozole,leucovorin calcium, leuprolide acetate, levamisole HCL, lidocaine,lomustine, maytansinoid, mechlorethamine HCL, medroxyprogesteroneacetate, megestrol acetate, melphalan HCL, mercaptipurine, mesna,methotrexate, methyltestosterone, mithramycin, mitomycin C, mitotane,mitoxantrone, nilutamide, octreotide acetate, ondansetron HCL,paclitaxel, pamidronate disodium, pentostatin, pilocarpine HCL,plimycin, polifeprosan 20 with carmustine implant, porfimer sodium,procaine, procarbazine HCL, propranolol, rituximab, sargramostim,streptozotocin, tamoxifen, taxol, teniposide, tenoposide, testolactone,tetracaine, thioepa chlorambucil, thioguanine, thiotepa, topotecan HCL,toremifene citrate, trastuzumab, tretinoin, valrubicin, vinblastinesulfate, vincristine sulfate, vinorelbine tartrate, bevacizumab,sutinib, sorafenib, 2-methoxyestradiol or 2ME2, finasunate, vatalanib,vandetanib, aflibercept, volociximab, etaracizumab (MEDI-522),cilengitide, erlotinib, cetuximab, panitumumab, gefitinib, trastuzumab,dovitinib, figitumumab, atacicept, rituximab, alemtuzumab, aldesleukine,atlizumab, tocilizumab, temsirolimus, everolimus, lucatumumab,dacetuzumab, HLL1, huN901-DM1, atiprimod, natalizumab, bortezomib,carfilzomib, marizomib, tanespimycin, saquinavir mesylate, ritonavir,nelfinavir mesylate, indinavir sulfate, belinostat, panobinostat,mapatumumab, lexatumumab, dulanermin, ABT-737, oblimersen, plitidepsin,talmapimod, P276-00, enzastaurin, tipifarnib, perifosine, imatinib,dasatinib, lenalidomide, thalidomide, simvastatin, and celecoxib.

In certain embodiments, a compound described herein is administered tothe subject prior to treatment with another chemotherapeutic agent,during treatment with another chemotherapeutic agent, afteradministration of another chemotherapeutic agent, or a combinationthereof.

In one aspect of the invention, a compound disclosed herein can bebeneficially administered in combination with any therapeutic regimenentailing radiotherapy, chemotherapy, or other therapeutic agents.

Neurological Disorders

Additional therapeutic uses of the compounds disclosed herein includeuse in the treatment and/or prevention of neurological disorders. Theserange from dementia disorders, such as Alzheimer's and Parkinson's wherecellular structure deformation is a common denominator; to diseases ofthe eye such as exfoliating glaucoma. In one embodiment provided hereinis a method of treating a neurological disorder, comprisingadministering an effective amount of a compound described herein. In oneembodiment, the neurological disorder is Alzheimer's disease. In oneembodiment, the neurological disorder is Parkinson's disease. In oneembodiment, the neurological disorder is amyotrophic lateral sclerosis.In one embodiment, the neurological disorder is central or peripheralnervous system damage, dysfunction, or complications stemming fromedema, injury, or trauma. In one embodiment, the neurological disorderis Multiple Sclerosis, Alzheimer's disease, Amyotrophic LateralSclerosis, Parkinson's disease, Huntington's disease, neuropathic pain,or spinal cord injury. In one embodiment of the invention, a compoundfor the treatment of the neurological disorder is selected from thecompounds of Formula I, II or III as described herein, or apharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof. In one non-limiting example, the neurological disorderis treated with Formula I(a), Formula I(b), Formula I(c), Formula I(d),Formula I(e), Formula I(f), Formula I(g), Formula I(h), Formula I(i),Formula I(j), Formula I(k), Formula I(l), Formula I(m), Formula I(n),Formula I(o), Formula I(p), Formula I(q), Formula I(r), Formula I(s),Formula I(t), Formula I(u), Formula I(v), Formula I(w), Formula I(x),Formula I(y), Formula I(z), Formula I(aa), Formula I(ab), Formula I(ac),Formula I(ad), Formula I(ae), Formula I(af), Formula I(ag), FormulaI(ah), Formula I(ai), Formula I(aj), Formula I(ak), Formula I(al),Formula I(am), Formula I(an), or Formula I(ao) as described herein, or apharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof.

According to this invention, the active ingredient is not provided as abotanical extract mixture or combination, but instead the activecompound is delivered in a highly pure form. In one embodiment, theinvention is a dosage form for the treatment of a neurologic disordersuch as Alzheimer's, Parkinson's, and dementia, wherein the activecompound has a purity of at least 96%, 97%, 98%, or 99%, without respectto fillers, stabilizers, or other inert or inactive ingredients. In analternative embodiment, the dosage form has two or more activeingredients, wherein only one of the active ingredients is selected fromcompounds of Formula I, II or III as described herein. In a furtheralternative embodiment, the dosage form has two or more activeingredients selected from compounds of Formula I, II or III as describedherein, wherein each active compound has a purity of at least 96%, 97%,98%, or 99%, without respect to fillers, stabilizers, or other inert orinactive ingredients. In a further alternative embodiment, the dosageform has two or more active ingredients selected from compounds ofFormula I, II or III as described herein, wherein each active compoundhas a purity of at least 96%, 97%, 98%, or 99%, without respect tofillers, stabilizers, or other inert or inactive ingredients.

Neurological Disorder Combination Therapies In one aspect of theinvention, the compounds disclosed herein can be beneficiallyadministered in combination with one or more additional therapeutics forbeneficial, additive or synergistic effect to treat a neurologicaldisorder. Therapeutic combinations can include the use of a compound asdescribed herein with a therapeutic used to treat a neurologicaldisorder.

Therapies used for the treatment of neurological disorders include, butare not limited to, Abstral, Aggrenox, Aggrenox, Ampyra, Amrix, Anexsia,Apokyn, Aptiom, ARICEPT, Avinza, Avonex, Axert, Axona, Banzel, Botox,Bromfenac, Butrans, Cambia, Carbaglu, Carbatrol, Cenestin, Cialis,Clonazepam, Comtan, Copaxone, Cuvposa, Cylert, Depakote, Durezol,Edluar, Eliquis, Embeda, Exalgo, Exelon, Exparel, Extavia, Fetzima,Focalin, Frova, Fycompa, Galzin, Gralise, Hetlioz, Horizant, Imitrex,Intermezzo, Intuniv, Invega, Iontocaine, Kadian, Kapvay, Levetiracetam,Lamictal, Lazanda, Levitra, Lidoderm Patch, Lunesta, Lupron Depot,Lusedra, Lyrica, Maxalt, Metadate CD, Migranal, Mirapex, Myobloc,Naltrexone HCl, Namenda, Neupro, Neurontin, NORCO tablets, Northera,Novantrone, Nucynta, Nuedexta, Nuvigil, Nymalize, Onfi, Onsolis, Oxecta,Oxtellar XR, Oxycodone and Aspirin, Poicor, Potiga, Pramipexole,Quadramet, Quillivant XR, Qutenza, Rebif, Redux, Relpax, Reminyl,Requip, Rilutek, Rozerem, Sabril, Selegiline, Silenor, Sonata, Sprix,Stavzor, Strattera, Subsys, Tasmar, Tegretol, Tivorbex, Topamax,Trileptal, Trokendi XR, Tysabri, Ultracet, UltraJect, VERSED, Viibryd,Vimpat, Vivitrol, Vpriv, Vyvanse, Xenazine, Xifaxan, Xyrem, Zanaflex,Zipsor, Zohydro ER, Zomig, Zonegran, and Zubsolv.

Fibrotic Disorders

In one embodiment provided herein is a method of treating and/orpreventing a fibrotic disorder, comprising administering an effectiveamount of a compound described herein.

In one embodiment, the fibrotic disorder is a surgical adhesion,osteoarthritis, or a visible skin scar, and rheumatoid arthritis. In oneembodiment provided herein is a method of treating a cardiovascularfibrotic disorder such as atherosclerosis or arteriosclerosis, liverfibrotic disorders, kidney fibrotic disorders, lung fibrotic disordersor periodontal fibrotic disorders, comprising administering to a subjectin need thereof a compound described herein. In additional embodiments,the compounds provided herein can be used to treat and/or preventdiseases entailing excess collagen and elastin deposition. In a furtherembodiment, the fibrotic disorder is selected from the group consistingof cutaneous keloid formation, progressive systemic sclerosis, livercirrhosis, idiopathic and pharmacologically induced pulmonary fibrosis,chronic graft-versus-host disease, scleroderma (local and systemic),Peyronie's disease, pharmacologically induced fibrosis of the penis,post-cystoscopic urethral stenosis, post-surgical internal adhesions,myelofibrosis, and idiopathic and pharmacologically inducedretroperitoneal fibrosis. In one embodiment of the invention, a compoundfor the treatment of a fibrotic disorder is selected from the compoundsof Formula I, II or III as described herein, or a pharmaceuticallyacceptable composition, salt, isotopic analog, or prodrug thereof. Inone non-limiting example, the fibrotic disorder is treated with FormulaI(a), Formula I(b), Formula I(c), Formula I(d), Formula I(e), FormulaI(f), Formula I(g), Formula I(h), Formula I(i), Formula I(j), FormulaI(k), Formula I(l), Formula I(m), Formula I(n), Formula I(o), FormulaI(p), Formula I(q), Formula I(r), Formula I(s), Formula I(t), FormulaI(u), Formula I(v), Formula I(w), Formula I(x), Formula I(y), FormulaI(z), Formula I(aa), Formula I(ab), Formula I(ac), Formula I(ad),Formula I(ae), Formula I(af), Formula I(ag), Formula I(ah), FormulaI(ai), Formula I(aj), Formula I(ak), Formula I(al), Formula I(am),Formula I(an), or Formula I(ao) as described herein, or apharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof.

According to this invention, the active ingredient is not provided as abotanical extract mixture or combination, but instead the activecompound is delivered in a highly pure form. In one embodiment, theinvention is a dosage form for the treatment of a fibrotic disorder,wherein the active compound has a purity of at least 96%, 97%, 98%, or99%, without respect to fillers, stabilizers, or other inert or inactiveingredients. In an alternative embodiment, the dosage form has two ormore active ingredients, wherein only one of the active ingredients isselected from compounds of Formula I, II or III as described herein. Ina further alternative embodiment, the dosage form has two or more activeingredients selected from compounds of Formula I, II or III as describedherein, wherein each active compound has a purity of at least 96%, 97%,98%, or 99%, without respect to fillers, stabilizers, or other inert orinactive ingredients.

Fibrotic Combination Therapies

In one aspect of the invention, the compounds disclosed herein can bebeneficially administered in combination with one or more therapeuticsused to treat fibrotic disorders in order to provide beneficial,additive or synergistic effect. Therapies used for the treatment offibrotic disorders include, but are not limited to, corticosteroids suchas prednisone, cyclophosamide (Cytoxan®), azathioprine (Imuran®),N-Acetylcysteine (NAC), and pirfenidone (Esbriet®, Pirfenex®, Pirespa®).

Pharmaceutical Compositions and Dosage Forms

In one aspect, the invention provides a pharmaceutical compositioncomprising a pharmaceutically effective amount of the compounds of thepresent invention and a pharmaceutically acceptable carrier.

The compounds provided herein are administered for medical therapy in atherapeutically effective amount. The amount of the compoundsadministered will typically be determined by a physician, in the lightof the relevant circumstances, including the condition to be treated,the chosen route of administration, the compound administered, the age,weight, and response of the individual patient, the severity of thepatient's symptoms, and the like.

The pharmaceutical compositions provided herein can be administered by avariety of routes including oral, parenteral, topical, rectal,subcutaneous, intravenous, intramuscular, and intranasal with apharmaceutical carrier suitable for such administration. In oneembodiment, the compounds are administered in a controlled releaseformulation.

The compositions for oral administration can take the form of bulkliquid solutions or suspensions, or bulk powders. Typically, thecompositions are presented in unit dosage forms to facilitate accuratedosing. The term “unit dosage forms” refers to physically discrete unitssuitable as unitary dosages for human subjects and other mammals, eachunit containing a predetermined quantity of active material calculatedto produce the desired therapeutic effect, in association with asuitable pharmaceutical excipient. Typical unit dosage forms includeprefilled, premeasured ampules or syringes of the liquid compositions orpills, tablets, capsules or the like in the case of solid compositions.In such compositions, the compound is usually a minor component (as anonlimiting example, from about 0.1 to about 50% by weight or preferablyfrom about 1 to about 40% by weight) with the remainder being variousvehicles or carriers and processing aids helpful for forming the desireddosing form. In one embodiment, the compound is present from about 1% toabout 10% by weight.

The compositions for topical administration can take the form of anemulsion, a cream, a lotion, a solution, an anhydrous stick, a serum,etc. The compositions can include from about 0.1% to about 50% by weightof the compound(s) of Formula I. In one embodiment the composition caninclude from about 0.1% to about 10% by weight of the compound(s) ofFormula I.

Liquid forms suitable for oral administration may include a suitableaqueous or nonaqueous vehicle with buffers, suspending and dispensingagents, colorants, flavors and the like. Solid forms may include, forexample, any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate; a glidant such as colloidal silicon dioxide; asweetening agent such as sucrose or saccharin; or a flavoring agent suchas peppermint, methyl salicylate, or orange flavoring.

Injectable compositions are typically based upon injectable sterilesaline or phosphate-buffered saline or other injectable carriers knownin the art.

The above-described components for orally administrable or injectablecompositions are merely representative. Other materials as well asprocessing techniques and the like are set forth in Part 8 ofRemington's Pharmaceutical Sciences, 17th edition, 1985, Mack PublishingCompany, Easton, Pa., which is incorporated herein by reference.

The compounds of this invention can also be administered in sustainedrelease forms or from sustained release drug delivery systems. Adescription of representative sustained release materials can be foundin Remington's Pharmaceutical Sciences.

In certain embodiments, the formulation comprises water. In anotherembodiment, the formulation comprises a cyclodextrin derivative. Incertain embodiments, the formulation compriseshexapropyl-β-cyclodextrin. In a more particular embodiment, theformulation comprises hexapropyl-β-cyclodextrin (10-50% in water).

The present invention also includes pharmaceutically acceptable acidaddition salts of compounds of the compounds of the invention. The acidswhich are used to prepare the pharmaceutically acceptable salts arethose which form non-toxic acid addition salts, i.e. salts containingpharmacologically acceptable anions such as the hydrochloride,hydroiodide, hydrobromide, nitrate, sulfate, bisulfate, phosphate,acetate, lactate, citrate, tartrate, succinate, maleate, fumarate,benzoate, para-toluenesulfonate, and the like.

Synthesis Preparation of Active Compounds:

The disclosed compounds can be made by the following general schemes.

In Scheme 1, Step 1, tall oil or safflower oil is crystallized from anorganic solvent such as acetone at reduced temperature to generatelinoleic acid. In Step 2, linoleic acid is directly reacted with thionylchloride in a combination of organic solvents such as dimethylformamideand cyclohexane at reduced temperature to generate an acid chloride. InStep 3, sodium omadine is directly reacted with bromotrichloromethane.The resulting complex is directly reacted with the acid chloride fromStep 2 to generate a bromodiene. In Step 4, lithiated furan is generatedby directly reacting furan with n-butyllithium. In Step 5, lithiatedn-butyllithium is directly reacted with the bromide from Step 3 togenerate a furan derivative.

In Scheme 2, Step 4, lithiated thiophene is produced by directlyreacting thiophene with n-butyllithium. In Step 5, lithiated thiopheneis directly reacted with the bromide of Step 3, Scheme 1 to produce athiophene derivative.

In Scheme 3, pyrrole is directly reacted with an acid in the presence ofzinc powder to generate a ketone. In Step 2, the ketone is directlyreacted with a reducing agent such as sodium borohydride in a proticsolvent such as 2-propanol to generate a pyrrole derivative.

In another embodiment, compounds of Formula I can be prepared bydirectly reacting furan or thiophene with an acid chloride using FriedelCraft conditions. The resulting ketone can be reduced with a reducingagent such as sodium borohydride in a protic solvent such as 2-propanol.Alternatively, the ketone can be directly reacted with ethanethiol togenerate a thioketal. The thioketal can be reduced using Raney nickel.

In another embodiment, compounds of Formula I can be prepared bydirectly reacting furaldehyde or thiophenaldehyde with a Wittig reagentto generate an alkene derivative.

In another embodiment, 2-(tri-n-butylstannyl)pyrrole and an acidchloride can be directly reacted using Stille coupling conditions, See,Mohamed, Y. M. A. and Hansen, T. V., Synthesis of mycalazol andmycalazal analogs with potent antiproliferating activities, Purr. Appl.Chem., 83:489-493, 2011. The resulting ketone can be reduced to thecorresponding alkane by directly reacting the ketone with palladium oncarbon under a hydrogen atmosphere in a protic solvent such as ethanoland an acid catalyst such as sulfuric acid.

EXAMPLES Example 1 Synthesis of 2-(8Z,11Z-heptadecadienyl)furan(Scheme 1) Step 1. Preparation of High Purity Linoleic Acid

A1) Obtain conjugated (“c”) and preferentially non-conjugated (“nc”)linoleic acid (C18) sourced from natural grapeseed (60% nc-10% c) orsafflower (60% nc-10% c).

B) Mix vegetable oil with purified water 2:1 to which has been added0.5% wt/wt of Candida rugosa enzymes (Amano 12K) or use an immobilizedenzyme bed and recirculate the water and oil mixture. In both examplesmaintain temperature at not more than 40° C. and not less than 35° C.under a nitrogen blanket. If enzymes are freely mixed then hold atmixing that allows complete recirculation of container contents every 60seconds and continue for 24 hours. Discontinue reaction and add heptaneor other non-polar solvent to solution (approx 0.5:1 ratio), stir anddecant under nitrogen and low light. Repeat three times to obtain thefatty acids. Evaporate solvent under nitrogen.

A2) Alternately and preferentially obtain Tall Oil fatty acids that arepredominately linoleic (75%) distributed as 90% non-conjugated and 10%conjugated.

C) Mix the fatty acids obtained in B or in A2 and mix with acetone in aratio of 1 part fatty acid to 3 parts acetone (weight:weight) and bringto −40° C. from 1 to 6 hours and hold for 48 hours at −40° C.

D) Vacuum filter using nominal 5-15 micron filter while chilled at lessthan −35° C. Evaporate solvent to recover very high purity linoleic acid(>95%).

E) Recover 99% pure linoleic acid (non-conjugated) with flashchromatography or critical fluids chromatography using standardseparation protocols.

Step 2. Preparation of Linoleoyl Chloride

A) Add 5 parts DMF (dimethyformamide) to 12,000 parts chilled (5° C.)cyclohexane and 3000 parts chilled (5° C.) linoleic acid and stir undera nitrogen flow of 0.2-0.4 L/min in an ice bath.

B) Add chilled (5° C.) thionyl chloride, SOCl₂, dropwise at 3 to 4mL/min.

C) After all of the thionyl chloride is added, allow the ice bath tocome to room temperature and continue stirring under nitrogen flow for24 hours or until all gas evolution has ceased.

D) Decant cyclohexane from tar layer and evaporate under nitrogen torecover clear amber liquid. Store well sealed in glass containers. Theproduct is obtained in a 90% yield.

Step 3. Preparation of Bromodiene

A) Mix C₅HSNOS (sodium omadine) and BrCCl₃ (bromotrichlormethane)together in a glass vessel and place at 5° C. for 24 hours to inducecrystal formation.

B) Add mixture from Step A to chilled, 5° C., cyclohexane in ice bathand begin nitrogen flow of 0.1-0.3 L/min.

C) Add dropwise chilled, 5° C., linoleoyl chloride from step 2) at notmore than 3-4 ml/min to reactor mixture in Step B.

D) After all linoleoyl chloride from step 2 has been added, continuestirring under nitrogen flow.

Allow ice bath to melt.

E) Note color change to yellow/red to indicate formation of bromodiene.

F) Follow with decanting and recovery of bromodiene using flashchromatography. The product was isolated in a 50% yield.

Step 4: Preparation Lithiated Furan

1) Prepare a 1M solution of butyllithium in hexane. Note: do not chillbelow 10° C. to prevent congealing.

2) Add chilled, 5° C., furan to chilled, 5° C., THF.

3) Maintain ice bath.

4) Add butyllithium in hexane dropwise at approximately 3-4 ml/min tochilled furan/THF solution.

5) Maintain ice bath and stir for not longer than 2 hours after allbutyllithium has been added.

The lithiated furan is obtained in a 90% yield.

Step 5: Synthesis of 2-(8Z,11Z-Heptadecadienyl)Furan

1. Add approximately 1 part cyclohexane and 1 part bromodiene from Step3 together and chill to 5° C. and begin nitrogen flow at approximately0.1 to 0.4 L per minute. Maintain temperature and stir.

2. Add chilled, 5° C., lithiated furan from Step 4 dropwise atapproximately 3-4 ml/min. Maintain temperature and stirring.

3. Note color change to dark reddish brown. After all lithiated furanhas been added maintain chilled reaction, nitrogen flow and stirring forone hour.

4. After one hour allow temperature to rise to ambient temperature andthen maintain stirring and nitrogen for 12 hours.

5. Wash product three times with 3% saline solution mixed withisopropanol or ethanol (1:1) and decant, evaporate solvent.

6. Purify with chromatography.

Example 2 Synthesis of 2-(8Z,11Z-heptadecadienyl)thiophene (Scheme 2)

Steps 1-3 are carried out as described in Example 1.

Step 4: Synthesis of Lithiated Thiophene

1) Prepare a 1M solution of butyllithium in hexane. Note: do not chillbelow 10° C. to prevent congealing.

2) Add chilled, 5° C., thiopene to chilled, 5° C., cyclohexane.

3) Maintain ice bath.

4) Add butyllithium in hexane dropwise at approximately 3-4 ml/min tochilled thiopene/cyclohexane solution.

5) Maintain ice bath and stir for not longer than 2 hours after allbutyllithium has been added.

6) Work up in standard manner to obtain product.

Step 5: Synthesis of 2-(8Z,11Z-heptadecadienyl)thiophene

1. Add approximately 1 part cyclohexane and 1 part bromodiene from Step3 together and chill to 5° C. and begin nitrogen flow at approximately0.1 to 0.4 L per minute. Maintain temperature and stir.

2. Add chilled, 5° C., lithiated thiophene from Step 4 dropwise atapproximately 3-4 ml/min.

Maintain temperature and stirring.

3. After all lithiated thiopene has been added maintain chilledreaction, nitrogen flow and stirring for one hour.

4. After one hour allow temperature to rise to ambient temperature andthen maintain stirring and nitrogen for 12 hours.

5. Wash product three times with 3% saline solution mixed withisopropanol or ethanol (1:1) and decant, evaporate solvent.

6. Purify with chromatography. The product was obtained in a 85% yield.

Example 3 Synthesis of 2-(8Z,11Z-heptadecadienyl)pyrrole (Scheme 3)

Step 1: A mixture of pyrrole (2.0 g, 29.8 mmol) 8Z,11Z-hexadeadienoicacid (17.83 g, 44.7 mmol) and zinc powder (3.88 g, 59.7 mmol) in toluene(75 ml) is stirred at room temperature until the pyrrole is no longerdetectable by thin layer chromatography or HPLC. The reaction isquenched with saturated sodium bicarbonate solution (50 ml) andextracted with ethyl acetate (3×30 ml). The combined organic layers arewashed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The product is purified by using silica gelcolumn chromatography that is eluted with a dichloromethane-methanolgradient.

Step 2:

To a stirred solution of the ketone from step 1 (1.63 g, 5.18 mmol), in150 ml of 2-propanol at ambient temperature is slowly added sodiumborohydride (1.34 g, 36.26 mmol). The reaction is heated at reflux andmonitored by thin layer chromatography or HPLC. Once the startingmaterial is no longer detected, the reaction is poured into 150 ml ofice-water and the solution is acidified with 10% aqueous HCl. Thereaction is extracted with dichloromethane (3×50 ml). The combinedorganic extracts are washed with water, brine, and dried over anhydrousNa₂SO₄, filtered and concentrated in vacuo. The product is purified byusing silica gel column chromatography that is eluted with adichloromethane-methanol gradient.

Example 4 2-(8Z,11Z-Heptadecadienyl)furan Formulation

According to one embodiment of the present invention,

A formulation is prepared comprising 2-(8Z,11Z-heptadecadienyl)furanhaving a purity of greater than 98% and a compound of Formula I having apurity of greater than 98% in a ratio of 80 parts to 20 parts. The twocomponents are added to a vegetable oil base and encapsulated for humanoral consumption. The total amount of 2-(8Z,11Z-heptadecadienyl)furanand a compound of Formula I as expressed as a percent of the finishedproduct can vary from 0.1 to 100%, with a preferred amount at about 5%.The vegetable oil carrier may be selected from any commonly produced forhuman consumption.

Example 5 Lipidic Furan Formulation

According to another embodiment of the invention,

The lipidic furan, 2-(8Z,11Z-heptadecadienyl)furan, can be incorporatedin a vegetable oil base for orally dosed pharmaceutical application. Inthis instance, it is preferable to provide a greater purity of thecompound of not less than 98% in order to ensure consistent andpredictable outcome of desired results and to take advantage of itsgeneral long term stability. The total amount of the compound asexpressed as a percent of the finished product can vary from 0.1 to100%, with a preferred amount at about 5% for better dietary uptake andabsorption in a vehicle of vegetable oil ordinarily provided for humanconsumption.

Example 6 Formula I(n) Inhibits Invasive Tumor Cell Migration

The effect of Formula I(n) on the migration of MDA-MB231 and Hs578Tinvasive breast tumor cells was tested using the Membrane InvasionCulture System (MICS) chamber. MDA-MB231 cells are breast adenocarcinomacells and are available from ATCC (Cailleau R, et al. Long-term humanbreast carcinoma cell lines of metastatic origin: preliminarycharacterization. In Vitro 14: 911-915, 1978). Hs578T cells are breastcarcinoma cells and are available from ATCC (Hackett A J, et al. Twosyngeneic cell lines from human breast tissue: the aneuploid mammaryepithelial (Hs 578T) and the diploid myoepithelial (Hs 578Bst) celllines. J. Natl. Cancer Inst. 58: 1795-1806, 1977).

The Membrane Invasion Culture System (MICS) chamber was assembled withCrosstex 10 μm polycarbonate membrane pre-soaked in gelatin for 12hours. The lower wells were fully loaded with DMEM supplemented with 10%FBS and 0, 5, 10 μl/10 ml Formula I(n) diluted in 0-20 μl/10 mlmethanol. The upper wells were loaded with 1 ml serum-freephenol-red-free media and corresponding Formula I(n)/methanol solution.100,000 cells/well were loaded and the chamber was incubated at 37° C.for 24 hours. After the incubation, the media from the upper part of thechamber was removed and non-migratory cells from the upper surface ofthe membrane were wiped away with Kimwipe. The cells were fixed onto themembrane by immersing it in 100% MeOH. Cells were stained in Eosinsolution for 25 seconds and in crystal violet solution for 35 seconds.The membrane was rinsed in water, placed onto a microscope slidepre-treated with immersion oil, covered with a coverslip and cells werecounted under a light microscope. Each sample was measured intriplicate.

As shown in FIG. 1, cells were treated with 5l or 10 μl/10 ml FormulaI(n) and evaluated compared to untreated control cells and control cellsexposed to the same concentration of methanol used as a solvent forFormula I(n) (10 μl/10 ml). As shown in FIG. 1, Formula I(n)significantly inhibited tumor cell migration in both MDA-MB231 andHs578T breast cancer cell lines.

Example 7 Formula I(n) Promotes a Stable Mesenchymal Cell Phenotype

Tumor cell behavior is affected to a great degree by the phenotype ofthe surrounding stromal cells and interactions between the epithelialand mesenchymal cell types. To determine if Formula I(n) also affectsmesenchymal cells, migratory ability of fibroblasts in the presence ofFormula I(n) was tested.

Confluent cultures of foreskin fibroblasts (Clonetics) were scratchedwith the tip of a 200 μl pipet tip, rinsed once with PBS and media waschanged to serum free media containing 0 (FIG. 2A, 2D, 2G) 5 (FIG. 2B,2E, 2H) or 10 (FIG. 2C, 2F) μl/10 ml Formula I(n). Pictures were takenat 0 (FIG. 2A, 2B, 2C), 12 (FIG. 2D, 2E, 2F) or 24 (FIG. 2G, 2H) hoursfollowing scratching and the distance between the migrating lips weremeasured in pixels. FIG. 3 is a graph of the distance between themigrating lips (as measured in pixels) vs. the time (hours) afteradministration of Formula I(n).

By 24 hours, the control (0 μl Formula I(n)) cells have completelyclosed the in vitro wound (FIG. 2G and FIG. 3). In contrast, FormulaI(n) treated cells showed reduced migration that for the higher FormulaI(n) concentration was almost completely diminished (FIG. 2E, 2F, 2H andFIG. 3).

Example 8 Formula I(n) Induces Cytoskeletal Reorganization and InhibitsTumor Cell Proliferation

Formula I(n) was next analyzed for its ability to induce cytoskeletalreorganization. Fibroblasts were treated at pre-confluent stages with 0(FIG. 4A), 5 μl (FIG. 4B), or 10 μl (FIG. 4C) Formula I(n) for 24 hoursand stained with phalloidin (actin-green) and propidium iodide(nuclei-red). Formula I(n) treated cells revealed a concentrationdependent increase of staining intensity of the actin cytoskeleton,numerous filopodia and reduced staining at adhesion sites compared tocontrols consistent with reduced cell migration (Seehttp://www.cellmigration.org/science/#overview for further discussion ofcell migration).

Confluent MDA-MB231 breast cancer cells were treated with 0 (FIG. 4.D),5 μl (E), or 10 μl (F) Formula I(n) for 24 hours and stained withphalloidin (actin-green) and propidium iodide (nuclei-red). PreconfluentMDA-MB231 breast cancer cells (FIG. 4) were treated with 0 (G), or 5 μl(H) μl Formula I(n) for 24 hours and stained with phalloidin(actin-green) and propidium iodide (nuclei-red).

As shown in FIG. 4, Formula I(n) treated tumor cells became elongated,spindle shaped, and consistent with their reduced ability to migrate,the actin filaments formed long parallel bundles in protrusions andspike-like thin sensory filopodia, but not broad lamellipodia that wouldbe strong foundations for cell movement. In addition, at both FormulaI(n) concentrations and at both stages of cell confluency, celldensities were significantly reduced upon treated with Formula I(n),thus reducing tumor cell proliferation.

Example 9 Reduced Focal Adhesion Kinase (FAK) [Tyr576] Phosphorylationin Formula I(n) Treated Cells

It has previously been shown that reduced migratory ability of variousinvasive breast tumor cells, including MDA-MB231, Hs578T (Payne S L,Fogelgren B, Hess A R, Seftor E A, Fong S F T, Csiszar K, Hendrix M J C,and Kirschmann D A. Lysyl oxidase regulates breast cancer cell migrationand adhesion through a hydrogen peroxide-mediated mechanism. Cancer Res.65:11429-36, 2005), and astrocytoma cells (Laczko R, Szauter K M, JansenM K, Hollosi P, Muranyi M, Molnar J, Fong K S K, Hinek A, Csiszar K.Active lysyl oxidase (LOX) correlates with FAK/paxillin activation andmigration in invasive astrocytes. Neuropathol Appl Neurobiol. 33:631-43,2007) was due to reduced activation of Focal Adhesion Kinase (FAK) anddiminished SRC and/or Paxillin signaling. In order to determine thestatus of FAK activation in Formula I(n) treated cells, thephosphorylation status of FAK[Tyr576] in Hs578T cells treated with 5 and10 μl Formula I(n) was analyzed.

In these experiments, Hs578T cells were cultured in standard DMEM, 1×Antibiotics and 10% FBS. Upon reaching confluency, cells were rinsedonce with PBS and media changed to serum free, phenol red free DMEM. Thecells were treated with 0, 5 or 10 μl Formula I(n)/10 ml media in 20 μlMeOH for 3 days. Protein from 6 days confluent conditioned cell culturemedia was collected and cell lysate samples were extracted using M-PERMammalian Protein Extraction Reagent (Pierce, Rockford, Ill., USA)supplemented with Halt protease inhibitor (Pierce) and phosphataseinhibitors Na₃VO₄ (10 mM) and NaF (160 mM). Following cell lysis,protein concentration with Bradford reagent was determined using aPolarstar Optima microplate reader (BMG Labtechnologies, Durham, N.C.,USA), and 20 μg protein was used for SDS-PAGE protein assay. The proteinsamples were size separated on 10% polyacrylamide gel and transferredonto PVDF membrane (Millipore, Billerica, Mass., USA) using semidryBio-Rad (Hercules, Calif., USA) transfer system. Ponceau staining of thePVDF membranes was performed to ensure equal loading. The membranes wereblocked with 5% non-fat dry milk in PBST overnight at 4° C. Membraneswere washed in PBST, and rabbit anti-FAK[pTyr576] (1:1000, BiosourceInc., Camarillo, Calif., USA) antibody was applied on the membrane for 1hour at room temperature. Membranes were then washed in PBST andincubated with anti-goat (Jackson Immuno Research, Baltimore, Pa., USA)horseradish peroxidase-conjugated secondary antibody forchemiluminescent detection. Subsequently, membranes were washed in PBSand incubated with ECL plus Western blotting reagent mix (AmershamPharmacia Biotech).

As shown in FIG. 5, Formula I(n) induced a concentration dependentdecrease of FAK[Tyr576] phosphorylation.

Example 10 Efficacy of the Compounds in Breast Tumors In Vivo

A HER2-driven model of breast cancer (Muller W J, Sinn E, Pattengale PK, Wallace R, Leder P. Single-step induction of mammary adenocarcinomain transgenic mice bearing the activated c-neu oncogene. Cell 1988; 54:105-15), that expresses c-neu (the mouse ortholog of human HER2) drivenby the MMTV promoter is used in the following example.

MMTV-neu mice are generated and observed post-lactation, with tumorsobserved with a median latency of approximately 25 weeks. Mice areenrolled in therapy studies when tumors reached a standard size (50-60mm3) that permit easy serial assessment. Tumor-bearing mice arecontinuously treated with an active compound added to their chow.MMTV-c-neu mice are examined weekly to assess tumor development bypalpation. Tumor volumes are calculated by the formula,Volume=[(width)²×length]/2. Tumor-bearing mice are euthanized at theindicated times due to predefined morbidity, tumor ulceration, or atumor size of more than 1.5 cm in diameter.

This specification has been described with reference to embodiments ofthe invention. The invention has been described with reference toassorted embodiments, which are illustrated by the accompanyingExamples. The invention can, however, be embodied in different forms andshould not be construed as limited to the embodiments set forth herein.Given the teaching herein, one of ordinary skill in the art will be ableto modify the invention for a desired purpose and such variations areconsidered within the scope of the invention.

What is claimed is:
 1. A method for the treatment of exfoliatingglaucoma in a human in need thereof, comprising administering to thehuman an effective amount of a compound of Formula I(t)


2. The method of claim 1, wherein the compound is suitable for topicaladministration.
 3. A method for the treatment of exfoliating glaucoma ina human in need thereof, comprising administering to the human aneffective amount of a compound of Formula I(u)

or a pharmaceutically acceptable salt thereof.
 4. The method of claim 3,wherein the compound is suitable for topical administration.
 5. A methodfor the treatment of exfoliating glaucoma in a human in need thereof,comprising administering to the human an effective amount of a compoundof Formula I(n)


6. The method of claim 5, wherein the compound is suitable for topicaladministration.